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The Experimental and Clinical Radiobiology Department was established as an independent research unit in 1997 as a result of a fusion between the Laboratory of Radiobiology from the Department of Radiotherapy, the Laboratory of DNA Topology, and a Clinical group working in the Radiotherapy Department. At the moment the Department of Experimental and Clinical Radiobiology consists of two laboratories, the Laboratory of Cell Radiobiology and the Laboratory of DNA Topology. Recently we are organizing the Laboratory of Clinical Proteomics. In past the Laboratory of Clinical Radiobiology was the part of our Department, which is currently affiliated at Department of Radiotherapy of our Institute.
The Department studies the influence of genotoxic factors which are applied in cancer therapy (ionizing radiation and antiancer drugs) on cells and on their genetic material, and the early and late effects of these therapies.
Main Topics of interest:
1. DNA damage and repair, studied on normal and tumor cells:
- the characterization of proteins which recognize DNA damage and participate in its repair;
- the influence of antioxidants on DNA damage induced by radiation;
2. The individual variability of radiosensitivity of cancer patients and healthy subjects, tested in vitro by genetic, cytogenetic and cellular tests.
3. The comparative assessment of the effectiveness of conventional, accelerated or continuous schemes of radiation dose fractionation:
- for radiotherapy of cancer patients;
- in multicellular megacolonies, an in vitro tumor model;
- with the use of mathematical modeling.
4. The identification of genes and proteins which are involved in cellular responses to radiation and modulate the radiation sensitivity of normal and malignant cells.
5. The regulation of molecular processes involved in terminal stages of apoptosis: DNA fragmentation and chromatin condensation.
The Department cooperates closely with the Institute of Automatics of the Silesian Technical University, Gliwice to develop mathematical models which help to understand the biological processes described above. Together, this Institute and the Experimental and Clinical Radiobiology Department conduct common scientific projects and organize conferences and seminars. Another close collaboration was established with several distinguished overseas laboratories, including Laval University in Quebec, Kanada (group of Prof. Ronald Hancock), UT Southwestern Medical Center in Dallas, USA (group of Prof. William Garrard), Rice University, Houston, USA (group of Prof. Mark Kimmel). We also have a close cooperation with the Silesian University in Katowice and Silesian Medical Academy.
The Head of the Experimental and Clinical Radiobiology Department coordinates several international exchange projects involving Ukraine, Belarus, Latvia and the Russian Federation. Many students perform pregraduate studies or experimental work for the Master's degree in the Radiobiology Department.
Since 1997 the Department of Experimental and Clinical Radiobiology Department organizes the "Gliwice Scientific Meeting", an annual Conferences with more than 100 Polish and international participants.
Head: Maria Wideł, PhD | 
Head: Joanna Rzeszowska - Wolny, PhD |
Head: Piotr Widłak, PhD | |
HEAD: Maria Wideł, PhD
Scientific staff:
HEAD: Joanna Rzeszowska - Wolny, PhD
Scientific staff:
HEAD: PIOTR WIDŁAK PhD
Scientific staff:
Main research interest:
Identification and functional analyses of proteins involved in cellular responses to ionizing radiation and other stress factors.
Currently we conduct projects related to:
- apoptotic nucleases (DFF40/CAD and EndoG) and mechanisms that regulate their activity;
- damaged DNA-binding (DDB) proteins involved in recognition of DNA damage;
- mechanisms of chromatin rearrangements in cells expose to genotoxic factors;
- serum proteomics and novel molecular markers of cancer.
Important works:
(click on a title shows abstract of publication)
Sikora E., Bielak-Żmijewska A., Magalska A., Piwocka K., Mosieniak G., Kalinowska M., Widłak P., Cymerman I.A., Bujnicki J.M.:Curcumin induces caspase-3-dependent apoptotic pathway but inhibits DNA fragmentation factor 40/caspase-activated DNase endonuclease in human Jurkat cells.
Curcumin is a natural pigment that has been shown to induce cell death in many cancer cells; however, the death mode depends on the cell type and curcumin concentration. Here we show that, in Jurkat cells, 50 micromol/L curcumin severely lowers cell survival and induces initial stage of chromatin condensation. It also induces caspase-3, which is sufficient to cleave DNA fragmentation factor 45 [DFF45/inhibitor of caspase-activated DNase (ICAD)], the inhibitor of DFF40/CAD endonuclease. However, the release of DFF40/CAD from its inhibitor does not lead to oligonucleosomal DNA degradation in curcumin-treated cells. Moreover, curcumin treatment protects cells from UVC-induced oligonucleosomal DNA degradation. In biochemical experiments using recombinant DFF activated with caspase-3, we show that curcumin inhibits plasmid DNA and chromatin degradation although it does not prevent activation of DFF40/CAD endonuclease after its release from the inhibitor. Using DNA-binding assay, we show that curcumin does not disrupt the DNA-DFF40/CAD interaction. Instead, molecular modeling indicates that the inhibitory effect of curcumin on DFF40/CAD activity results from curcumin binding to the active center of DFF40/CAD endonuclease. [J Cell Biochem. 2005 Apr 15;94(6):1078-87. Review]
Widłak P., Garrard W.T.:The apoptotic endonuclease DFF40/CAD is inhibited by RNA, heparin and other polyanions.
DFF40/CAD, the major apoptotic nuclease, is specific for double-stranded DNA. However, RNA and single-stranded DNA, though not substrates for the enzyme, compete with double-stranded DNA and inhibit its cleavage by the nuclease. In addition, other anionic polymers, like poly-glutamic acid and heparin also inhibit DFF40/CAD, the latter one being highly effective at nanomolar concentrations. The inhibitory poly-anions bind to the nuclease and impair its ability to bind double-stranded DNA. We propose that such poly-anions bind to the positively charged surface formed by alpha4 helices of the DFF40/CAD homodimer. This surface has been proposed recently to bind to either the major groove of DNA or poly (ADP-ribose), another inhibitor of the nuclease. [Apoptosis. 2006 Aug;11(8):1331-7]
Widłak P., Garrard W.T.:Unique features of the apoptotic endonuclease DFF40/CAD relative to micrococcal nuclease as a structural probe for chromatin.
The gold standard for studies of nucleosomal chromatin structure for the past 30 years has been the enzyme micrococcal nuclease (MNase). During the course of our studies on the elucidation of the mechanism of action of the apoptotic nuclease DNA fragmentation factor-40 / caspase-activated deoxyribonuclease (DFF40/CAD) on naked DNA and chromatin substrates, it became clear that this enzyme is superior in certain respects to MNase for studying several aspects of chromatin structure. Here we review our published results supporting this statement. Relative to MNase, we have found that DFF40/CAD has the following properties: (i) it does not cut within nucleosomes to generate subnucleosomal DNA fragments; (ii) it is more specific for the linker regions between nucleosomes; (iii) it lacks exonuclease activity; (iv) it is specific for double-stranded DNA and makes exclusively double-stranded breaks; and (v) it attacks histone-H1-containing chromatin more efficiently. Taken together, these facts explain why DFF40/CAD generates sharper oligonucleosomal DNA ladders compared with those generated by MNase. We therefore recommend the following uses for DFF40/CAD for chromatin research: nucleosome isolation, chromatin-remodeling assays, repeat length measurements, and nucleosome-positioning assays along specific sequences. Other uses include footprinting assays of transcription factor positions, shearing chromatin for immunopreciptitation experiments (ChIP), and shearing DNA for recombinant DNA library preparation or for shotgun cloning for sequencing. [Biochem Cell Biol. 2006 Aug;84(4):405-10]
Widłak P., Pietrowska M., Łanuszewska J.:The role of chromatin proteins in DNA damage recognition and repair.
The structure of chromatin is the major factor determining the rate and efficiency of DNA repair. Chromatin remodeling events such as rearrangement of nucleosomes and higher order chromatin structures are indispensable features of repair processes. During the last decade numerous chromatin proteins have been identified that preferentially bind to different types of DNA damage. The HMGB proteins, which preferentially interact with DNA intrastrand crosslinks induced by cisplatin, are the archetypal example of such proteins. Several hypothetical models have been proposed describing the role of such damage-binding chromatin proteins. The damage shielding model postulates that binding of chromatin proteins to damaged DNA might disturb damage recognition by repair factors and impair its removal. Alternatively, the damage-recognition/signaling model proposes that the binding of specific chromatin proteins to damaged DNA could serve as a hallmark to be recognized by repair proteins. Additionally, the binding of specific chromatin proteins to damaged DNA could induce chromatin remodeling at the damage site and indirectly affect its repair. This paper aims to critically review current experimental data in relation to such possible roles of chromatin proteins.[Histochem Cell Biol. 2006 Jan;125(1-2):119-26. Review]
Widłak P., Garrard W.T.:Discovery, Regulation and Action of the Major Apoptotic Nucleases DFF40/CAD and Endonuclease G.
Toward the end of the twentieth and beginning of the twenty-first centuries clever in vitro biochemical complementation experiments and genetic screens from the laboratories of Xiaodong Wang, Shigekazu Nagata and Ding Xue led to the discovery of two major apoptotic nucleases, termed DNA fragmentation factor (DFF) or caspase-activated DNase (CAD) and endonuclease G (Endo G). Both endonucleases attack chromatin to yield 3-hydroxyl groups and 5-phosphate residues, first at the level of 50-300 kb cleavage products and next at the level of internucleosomal DNA fragmentation, but these nucleases possess completely different cellular locations in normal cells and are regulated in vastly different ways. In non-apoptotic cells, DFF exists in the nucleus as a heterodimer, composed of a 45 kD chaperone and inhibitor subunit (DFF45) [also called inhibitor of CAD (ICAD-L)] and a 40 kD latent nuclease subunit (DFF40/CAD). Apoptotic activation of caspase-3 or 7 results in the cleavage of DFF45/ICAD and release of active DFF40/CAD nuclease. DFF40s nuclease activity is further activated by specific chromosomal proteins, such as histone H1, HMGB1/2 and topoisomerase II. DFF is regulated by multiple pre- and post-activation fail-safe steps, which include the requirements for DFF45/ICAD, Hsp70 and Hsp40 proteins to mediate appropriate folding during translation to generate a potentially activatable nuclease, and the synthesis in stoichiometric excess of the inhibitors (DFF45/35)(ICAD-S/L). By contrast, endonuclease G resides in the mitochondrial intermembrane space in normal cells, and is released into the nucleus upon apoptotic disruption of mitochondrial membrane permeability in association with co-activators such as apoptosis-inducing factor (AIF). Understanding further regulatory check-points involved in safeguarding non-apoptotic cells against accidental activation of these nucleases remain as future challenges, as well as designing ways to selectively activate these nucleases in tumor cells. [J. Cell. Biochem. (2005) in press]
Widłak P., Kalinowska M., Parseghian M.H., Lu X., Hansen J.C., Garrard W.T.: The histone H1 C-terminal domain binds to the apoptotic nuclease, DNA Fragmentation Factor (DFF40/CAD) and stimulates DNA cleavage.
The apoptotic nuclease, DNA fragmentation factor (DFF40/CAD), is primarily responsible for internucleosomal DNA cleavage during the terminal stages of programmed cell death. Previously we demonstrated that histone H1 greatly stimulates naked DNA cleavage by this nuclease. Here we investigate the mechanism of this stimulation with native and recombinant mouse and human histone H1 species. Using a series of truncation mutants of recombinant histone H1-0, we demonstrate that the H1 C-terminal domain (CTD) is responsible for activation of DFF40/CAD. We show further that the intact histone H1-0 CTD and certain synthetic CTD fragments bind to DFF40/CAD, and confer upon it an increased ability to bind to DNA. Interestingly, we find that each of the six somatic cell histone H1 isoforms, whose CTDs differ significantly in primary sequence but not amino acid composition, equally activate DFF40/CAD. We conclude that the interactions identified here between the histone H1 CTD and DFF40/CAD target and activate linker DNA cleavage during the terminal stages of apoptosis. [Biochemistry, (2005) in press]
Kalinowska M., Garncarz W., Pietrowska M., Garrard W.T., Widłak P.: Regulation of the human apoptotic DNase/RNase Endonuclease G: involvement of Hsp70 and ATP.
Endonuclease G (EndoG) is a mitochondrial enzyme that becomes an apoptotic nuclease when released from the mitochondrial intermembrane space. EndoG will digest either DNA or RNA, but at physiological ionic strength, RNA is a much more favorable substrate as compared to chromatin. This indicates that EndoGs major in vivo function(s) may be: (i) an apoptotic RNase, and/or (ii) an apoptotic DNase in the presence of additional co-activators. In the present study we have searched for factors that modulate the activity of human EndoG on DNA substrates. We demonstrate that EndoG forms complexes with AIF and FEN-1 but not with PCNA. Interestingly, heat shock proteins 70 interact with EndoG and are involved in the regulation of its activity. Purified Hsp70 prevented stimulation of EndoG DNase activity by other nuclear factors in the ATP-dependent manner. [Apoptosis (2005) in press]
Pietrowska M., Widłak P.: Characterization of a novel protein that specifically binds to DNA modified by N-acetoxy-acetylaminofluorene and cis-diamminedichloroplatinum.
Proteins recognizing DNA damaged by the chemical carcinogen N-acetoxy-acetylaminofluorene (AAAF) were analyzed in nuclear extracts from rat tissues, using a 36 bp oligonucleotide as substrate and electrophoretic mobility shift and Southwestern blot assays. One major damage-recognition protein was detected, which amount was estimated as at least 105 copies per cell. Levels of this protein were similar in extracts from brain, kidney and liver, but much lower in extracts from testis. Affinity of detected protein for DNA damaged by AAAF was ~70-fold higher than for undamaged DNA. DNA damaged by cis-diamminedichloroplatinum (cis-DDP), benzo(a)pyrene diolepoxide (BPDE) or UV-radiation also binds this protein with increased affinity, the former more strongly and the latter two more weakly as compared to AAAF-damaged DNA. Detected AAAF/DDP-damaged DNA binding (AAAF/DDP-DDB) protein was distinct from histone H1 or HMGB proteins, which were previously known to posses high affinity for cis-DDP-damaged DNA, and had a molecular mass of about 25 kDa. The level of this damage-recognition protein was not affected in rats treated with the carcinogen 2-acetylaminofluorene. The activity of AAAF/DDP-DDB protein could be detected also in extracts from mouse liver cells but not from Hep2G human hepatocellular carcinoma. [Acta Biochim. Polon. 52 (2005) in press]
Widłak P.: DNA microarrays, a novel approach in studies of chromatin structure.
The DNA microarray technology delivers an experimental tool that allows surveying expression of genetic information on a genome-wide scale at the level of single genes - for the new field termed functional genomics. Gene expression profiling - the primary application of DNA microarrays technology - generates monumental amounts of information concerning the functioning of genes, cells and organisms. However, the expression of genetic information is regulated by a number of factors that cannot be directly targeted by standard gene expression profiling. The genetic material of eukaryotic cells is packed into chromatin which provides the compaction and organization of DNA for replication, repair and recombination processes, and is the major epigenetic factor determining the expression of genetic information. Genomic DNA can be methylated and this modification modulates interactions with proteins which change the functional status of genes. Both chromatin structure and transcriptional activity are affected by the processes of replication, recombination and repair. Modified DNA microarray technology could be applied to genome-wide study of epigenetic factors and processes that modulate the expression of genetic information. Attempts to use DNA microarrays in studies of chromatin packing state, chromatin/DNA-binding protein distribution and DNA methylation pattern on a genome-wide scale are briefly reviewed in this paper. [Acta Biochim. Polon. 51 (2004) 1-8].
Weil M.R., Widłak P., Minna J.D, Garner H.R.: Global survey of chromatin accessibility using DNA microarrays.
An increasing number of studies indicate a central role for chromatin remodeling in the regulation of gene expression. Current methods for high-resolution studies of the relationship between chromatin accessibility and transcription are low throughput, making a genome-wide study impractical. To enable the simultaneous measurement of the global chromatin accessibility state at the resolution of single genes, we developed the Chromatin Array technique, in which chromatin is separated by its condensation state using either the solubility differences of mono- and oligonucleosomes in specific buffers or controlled DNase I digestion and selection of the large refractory (condensed) DNA fragments. By probing with a comparative genomic hybridization style microarray, we can determine the condensation state of thousands of individual loci and correlate this with transcriptional activity. Applying this technique to the breast tumor model cell line, MCF7, we found that when the condensation is homogeneous in the population of cells, expression is inversely proportional to the level of accessibility and the two methods of accessibility-based target selection correlate well. Using functional annotation and comparative genomic hybridization data, we have begun to decipher the possible biological implications of the relationship between chromatin accessibility and expression. [Genome Res. 14 (2004) 1374-1381]
Łanuszewska J., Widłak P.: The truncation of Ku86 in human lymphocytes.
The Ku heterodimer, which consists of Ku70 and Ku86 subunits, is a major sensor of DNA breaks. A truncated form of Ku86 lacking its C-terminus, termed Ku86 variant, has been detected in extracts from different human cells. Here we report that in human lymphocytes the Ku86 variant is not present in vivo but is generated in vitro upon cell lysis by a trypsin-like protease. The resulting Ku86 variant exists exclusively in complexes with Ku70, which possess strong affinity to DNA double strand termini. In different blood donors the levels of Ku86 variant correlated with the magnitude of radiation induced DNA breaks. [Cancer Lett. 205 (2004) 197-205]
Horak S., Polańska J., Widłak P.:Bulky DNA adducts in human sperm: relationship with fertility, semen quality, smoking, and environmental factors.
The integrity of DNA of spermatogenic cells can be affected by endogenous and exogenous genotoxic factors. Resulting DNA damage in spermatozoa may significantly contribute to impaired fertility. Here, the 32P-postlabeling method was used to analyze the levels of bulky DNA adducts in sperm cells in a group of 179 males, either healthy donors or patients with an impaired fertility. When all donors were analyzed, the levels of bulky DNA adducts was 1.2-fold higher in smokers than in non-smokers, but the difference was not statistically significant (P = 0.054). However, a statistically significant difference existed between current smokers and never smokers among the healthy individuals (1.7-fold increase, P = 0.008). No correlation between alcohol or coffee consumption and sperm DNA adducts was found. The levels of DNA adducts in sperm seemed to be unaffected by environmental and occupational factors. On the other hand, groups of healthy persons and patients with male-factor infertility differed significantly with respect to the level of bulky DNA adducts (P = 0.012). A significant negative correlation between DNA adducts and sperm concentration or sperm motility existed among patients with an impaired fertility (n = 93; P < 0.029, rS = -0.225). These results suggest that DNA adducts in sperm cells can be applied as potential biomarkers in studies of human infertility. [Mutation Res. 537 (2003) 53-65]
Widłak P., Łanuszewska J., Cary R. B., Garrard W. T.: Subunit structures and stoichiometries of human DFF proteins before and after induction of apoptosis.
DNA fragmentation factor (DFF) is one of the major endonucleases responsible for internucleosomal DNA cleavage during apoptosis. Understanding the regulatory checkpoints involved in safeguarding non-apoptotic cells against accidental activation of this nuclease is as important as elucidating its activation mechanisms during apoptosis. Here we address these issues by determining DFF native subunit structures and stoichiometries in human cells before and after induction of apoptosis using the technique of native pore-exclusion limit electrophoresis in combination with Western analyses. For comparison, we employed similar techniques with recombinant proteins in conjunction with atomic force microscopy. Before induction of apoptosis, the expression of DFF subunits varied widely among the cell types studied, and the chaperone/inhibitor subunits DFF45 and DFF35 unexpectedly existed primarily as monomers in vast excess of the latent nuclease subunit, DFF40, which was stoichiometrically associated with DFF45 to form heterodimers. DFF35 was exclusively cytoplasmic as a monomer. Nuclease activation upon caspase-3 cleavage of DFF45/DFF35 was accompanied by DFF40 homo-oligomer formation, with a tetramer being the smallest unit. Interestingly, intact DFF45 can inhibit nuclease activity by associating with these homo-oligomers without mediating their disassembly. We conclude that DFF nuclease is regulated by multiple pre- and post-activation fail-safe steps. [J. Biol. Chem. 278 (2003) 26915-26922]
Widłak P., Fujarewicz K.: The analysis of chromatin condensation state and transcriptional activity using DNA microarrays.
The DNA microarray-based technique has been developed to semi-quantitatively measure the in vivo global chromatin condensation state at the resolution of a gene. Chromatin was fractionated due to the differential solubility of histone H1-containing and histone H1-free nucleosomes. A set of genes non-randomly distributed between histone H1-free (uncondensed or open) and histone H1-containing (condensed or closed) chromatin fractions has been identified. The transcript levels have been measured for the same group of genes. The correlation between transcriptional activity and chromatin fraction distribution of particular genes has been established. [J. Med. Inf. Technol. 6 (2003) IP13-IP19]
Widłak P., Palyvoda O., Kumala S., Garrard W.T.: Modeling apoptotic chromatin condensation in normal cell nuclei.
Hallmarks of the terminal stages of apoptosis are genomic DNA fragmentation and chromatin condensation. Here, we have studied the mechanism of condensation both in vitro and in vivo. We found that DNA fragmentation per se of isolated nuclei from non-apoptotic cells induced chromatin condensation that closely resembles the morphology seen in apoptotic cells, independent of ATP utilization, at physiological ionic strengths. Interestingly, chromatin condensation was accompanied by release of nuclear actin, and both condensation and actin release could be blocked by reversibly pretreating nuclei with Ca, Cu, diamide, or low pH, procedures shown to stabilize internal nuclear components. Moreover, specific inhibition of nuclear F-actin depolymerization or promotion of its formation also reduced chromatin condensation. Chromatin condensation could also be inhibited by exposing nuclei to reagents that bind to the DNA minor groove, disrupting native nucleosomal DNA wrapping. In addition, in cultured cells undergoing apoptosis, drugs that inhibit depolymerization of actin or bind to the minor groove also reduced chromatin condensation, but not DNA fragmentation. Therefore, the ability of chromatin fragments with intact nucleosomes to form large clumps of condensed chromatin during apoptosis requires the apparent disassembly of internal nuclear structures that may normally constrain chromosome subdomains in nonapoptotic cells. [J. Biol. Chem. 277 (2002) 21683-21690]
Widłak P., Li L.Y., Wang X., Garrard W.T.: Action of recombinant human apoptotic endonuclease G on naked DNA and chromatin substrates; cooperation with exonuclease and DNaseI.
Endonuclease G (endoG) is released from mitochondria during apoptosis and is in part responsible for internucleosomal DNA cleavage. Here we report the action of the purified human recombinant form of this endonuclease on naked DNA and chromatin substrates. The addition of the protein to isolated nuclei from nonapoptotic cells first induces higher order chromatin cleavage into DNA fragments > 50 kb in length, followed by inter- and intranucleosomal DNA cleavages with products possessing significant internal single-stranded nicks spaced at nucleosomal (~190 bases) and subnucleosomal (~10 bases) periodicities. We demonstrate that both exonucleases and DNase I stimulate the ability of endoG to generate double-stranded DNA cleavage products at physiological ionic strengths, suggesting that these activities work in concert with endoG in apoptotic cells to ensure efficient DNA breakdown. [J. Biol. Chem., 276 (2001) 4840448409]
Widłak P., Garrard W.T.:Ionic and cofactor requirements for the activity of the apoptotic endonuclease DFF40/CAD.
The endonuclease DFF40/CAD mediates regulated DNA fragmentation and chromatin condensation in cells undergoing apoptosis. Here we report the enzyme's co-factor requirements, and demonstrate that the ionic changes that occur in apoptotic cells maximize DFF40/CAD activity. The nuclease requires Mg2+, exhibits a trace of activity in the presence of Mn2+, is not co-stimulated by Ca2+, is inhibited by Zn2+ or Cu2+, and has high activity over a rather broad pH range (7.0-8.5). The enzyme is thermally unstable, and is rapidly inactivated at 42 degrees C. Enzyme activity is markedly affected by ionic strength. At the optimal [K+] of 50-125 mM, which is in the range of the cytoplasmic [K+] for cells undergoing apoptosis, the activity of DFF40/CAD for naked DNA cleavage is about 100-fold higher than at 0 or 200 mM [K+]. Although these ranges of ionic strength do not affect DFF40 homo-oligomer formation, at higher ionic strengths the enzyme introduces single-stranded nicks into supercoiled DNA. [Mol. Cell. Biochem. 218 (2001) 125-130]
Widłak P.: DFF40/CAD hypersensitive sites are potentially involved in high molecular weight DNA fragmentation during apoptosis.
Sequential cleavage of genomic DNA into large-scale DNA fragments of 50-300-kb, followed by formation of mono- and oligonucleosomal DNA fragments, is a biochemical hallmark of programmed cell death (apoptosis). The endonuclease DFF40/CAD mediates regulated internucleosomal DNA fragmentation and chromatin condensation in cells undergoing apoptosis. DFF40 hypersensitive sites were detected in purified HeLa cell nuclei, and excision of 50-kb DNA fragments preceded formation of oligonucleosomal DNA ladders in nuclei treated with the nuclease. Topoisomerase II, but not topoisomerase I, stimulates DFF40 activity on plasmid DNA substrates. This suggests that interactions of DFF with the nuclear matrix-bound topoisomerase II may be involved in formation of DFF40 hypersensitive sites. [Cell. Mol. Biol. Lett. 5 (2000) 373-379]
Widłak P., Li P., Wang X., Garrard W.T.: Cleavage preferences of the apoptotic endonuclease DFF40 (Caspase-activated DNase or Nuclease) on naked DNA and chromatin substrates.
Here we report the co-factor requirements for DNA fragmentation factor (DFF) endonuclease and characterize its cleavage sites on naked DNA and chromatin substrates. The endonuclease exhibits a pH optimum of 7.5, requires Mg2+, not Ca2+, and is inhibited by Zn2+. The enzyme generates blunt ends or ends with 1-base 5-overhangs possessing 5-phosphate and 3-hydroxyl groups and is specific for double- and not single-stranded DNA or RNA. DFF endonuclease has a moderately greater sequence preference than micrococcal nuclease or DNase I, and the sites attacked possess a dyad axis of symmetry with respect to purine and pyrimidine content. Using HeLa cell nuclei or chromatin reconstituted on a 5 S rRNA gene tandem array, we prove that the enzyme attacks chromatin in the internucleosomal linker, generating oligonucleosomal DNA ladders sharper than those created by micrococcal nuclease. Histone H1, high mobility group-1, and topoisomerase II activate DFF endonuclease activity on naked DNA substrates but much less so on chromatin substrates. We conclude that DFF is a useful reagent for chromatin research. [J. Biol. Chem. 275 (2000) 8226-8232]
Łanuszewska J., Widłak P.: High mobility group 1 and 2 proteins bind preferentially to DNA that contains bulky adducts induced by benzo[a]pyrene diol epoxide and N-acetoxy-acetyl-aminofluorene.
High mobility group (HMG) proteins 1 and 2 are abundant non-histone chromosomal proteins that bind preferentially DNA that is bent or underwound. Previous studies have shown that these proteins preferentially bind to DNA damaged by the crosslinking agents cis-diammine-dichloro-platinum(II), chromium(III) and UV-C radiation. Here we have studied the binding of HMG-1/2 proteins to a duplex oligonucleotide damaged by benzo(a)pyrene diol epoxide or N-acetoxy-acetylaminofuorene using an electrophoretic mobility shift assay. Both chemicals induce monoadducts that are known to distort DNA structure. The affinities of HMG-1/2 for DNA damaged by benzo[a]pyrene diol epoxide or N-acetoxy-acetylaminofuorene were similar to that for UV-irradiated DNA, which were an order of magnitude higher than for undamaged DNA. In contrast, DNA modified by dimethyl sulfate was not preferentially recognised by HMG-1/2. [Cancer Lett. 158 (2000) 17-25]
Pietrowska M., Łanuszewska J., Walter Z., Rzeszowska-Wolny J., Widłak P.:Detection and characterization of rat protein recognizing DNA damaged by N-acetoxy-acetyl-aminofluorene.
Proteins recognizing DNA damaged by the chemical carcinogen N-acetoxy-acetyl-aminofluorene (AAAF) were analyzed in nuclear extracts from rat tissues, using a 36 bp oligonucleotide as the substrate and an electrophoretic mobility shift assay. Two major proteins that form complexes with DNA damaged by AAAF were detected; one of them also bound DNA damaged by cis-diammine-dichloroplatinum. The complex specific for AAAF-damaged DNA contained protein loosely attached to nuclear components. It was extracted with 0.1 M NaCl. The amount of this protein was estimated at about 105 copies per liver cell nucleus, and its probable size was about 42 kDa as detected by the Southwestern blotting assay. Its affinity for DNA damaged by AAAF was ~10-fold higher than that for undamaged DNA. Analogous AAAF-DDB (damaged-DNA-binding) proteins were also detected in extracts from rat brain, testis and kidney tissue. The levels of such proteins were not affected in rats treated with the carcinogen 2-acetylaminofluorene. [Cell. Mol. Biol. Lett. 5 (2000) 423-431]
Research grants - current and completed:
Grant KBN 3 P05A 104 24 (01.04.2003 - 04.05.2006)
Factors that regulate the activities of the apoptotic nucleases: DFF40/CAD and endonuclease G.
Grant KBN 3 P05A 105 24 (01.04.2003 - 01.05.2005)
Chromatin protein that recognizes changes in DNA structure induced by N-acetoxy-acetylaminofluorene i cisplatine.
Grant KBN 6 P04A 030 20 (01.06.2001 - 30.05.2002)
Rat liver chromatin protein that binds to DNA damaged by N-acetoxy-acetyloaminofluorene.
Grant KBN 6 P04A 013 17 (01.10.1999 - 30.09.2002)
Catalytical properties of DFF4/CAD, the apoptotic endonuclease.
Grant KBN 6 P04A 041 12 (01.05.1997 - 30.04.1999)
The role of damaged DNA-binding proteins in repair of chemically induced DNA damage.
double clik on a year shows a list of publications
2007:
- Widłak W., Vydra N., Malusecka E., Dudaladava V., Winiarski B.,Scieglinska D., Widłak P. (2007): Heat shock transcription factor 1 down-regulates spermatocyte-specific 70 kDa heat shock protein expression prior to the induction of apoptosis in mouse testes. Genes Cells 12: 487-499.
- Xiao F., Widłak P., Garrard W.T. (2007): Engineered apoptotic nucleases for chromatin research. Nucl. Acids Res. 35, e93
- Horak S., Olejek A., Widłak P. (2007): Sperm DNA adducts impair fertilization during ICSI but not during IVF. Folia Histochem. Cytobiol. 45, suppl. 1: 1-6.
- Ryabokon N.I., Goncharova R.I., Duburs G., Hancock R., Rzeszowska-Wolny J. Changes in poly(ADP-ribose) level modulate the kinetics of DNA strand break rejoining.
Mutat Res. 2008 Jan 1;637 (1-2): 173-81
2006:
- Konopacka M., Rzeszowska-Wolny J. (2006): The bystander effect-induced formation of micronucleated cells is inhibited by antioxidants, but the parallel induction of apoptosis and loss of viability are not affected. Mutat Res. 593, 32-38.
- Kramer-Marek G., Serpa C., Szurko A., Wideł M., Sochanik A., Snietura M., Kus P., Nunes R.M., Arnaut L.G., Ratuszna A. (2006): Spectroscopic properties and photodynamic effects of new lipophilic porphyrin derivatives: efficacy, localisation and cell death pathways. J. Photochem. Photobiol. 84: 1-14.
- Liu Z., Widłak P., Zou Y., Xiao F., Oh M., Li S., Chang M.Y., Shay J. W., Garrard W.T. (2006): A recombination silencer that specifies heterochromatin positioning and Ikaros association in the immunoglobulin kappa locus. Immunity 24, 405-415.
- Przybyszewski W.M., Wideł M., Rzeszowska-Wolny J. (2006): Cardiotoxic consequences of ionizing radiation and anthracyclines. Post. Hig. Med. Dosw. 60, 397-405.
- Sikora E., Bielak-Żmijewska A., Magalska A., Piwocka K., Mosieniak G., Kalinowska M., Widłak P., Cymerman I.A., Bujnicki J.M. (2006): Curcumin induces caspase-3-dependent apoptotic pathway but inhibits DNA fragmentation factor 40/caspase-activated DNase endonuclease in human Jurkat cells. Mol. Cancer Ther. 5, 927-934.
- Wideł M.S., Wideł M. (2006): Mechanisms of metastasis and molecular markers of malignant tumor progression. I. Colorectal cancer. Post. Hig. Med. Dosw. 60, 453-470.
- Widłak P., Garrard W.T. (2006): The apoptotic endonuclease DFF40/CAD is inhibited by RNA, heparin and other polyanions. Apoptosis 11, 1331-1337.
- Widłak P., Garrard W.T. (2006): Unique features of the apoptotic endonuclease DFF40/CAD relative to micrococcal nuclease as a structural probe for chromatin. Biochem. Cell Biol. 84, 405-410.
- Widłak P., Pietrowska M., Łanuszewska J. (2006): The role of chromatin proteins in DNA damage recognition and repair. Histochem Cell Biol. 125, 119-126.
- Vydra N., Małusecka E., Jarząb M., Lisowska K., Głowala-Kosińska M., Benedykt K., Widłak P., Krawczyk Z., Widłak W. (2006): Spermatocyte-specific expression of constitutively active heat shock factor 1 induces HSP70i-resistant apoptosis in male germ cells. Cell Death Differ. 13, 212-222.
2005:
- Kalinowska M., Garncarz W., Pietrowska M., Garrard W.T., Widłak P. (2005):Regulation of the human apoptotic DNase/RNase EndoG; involvement of Hsp70 and ATP. Apoptosis 10, 821-830.
- Pietrowska M., Widłak P. (2005): Characterization of a novel protein that specifically binds to DNA modified by N-acetoxy-acetylaminofluorene and cis-diamminedichloroplatinum. Acta Biochim. Polon., 52, 867-874.
- Przybyszewski W., Kasperczyk J., Stokłosa K., Bkhiyan A. (2005):Uszkodzenia DNA powodowane przez produkty peroksydacji lipidów. Post. Hig. Med. Dośw.59: 75-81.
- Przybyszewski W., Wideł M., Szurko A., Rzeszowska- Wolny J., Polaniak R., Matulewicz Ł., Maniakowski Z., Birkner E. (2005): Contrasting effects of low vs. high dose-radiation on lipid peroxidation DNA damage, and antioxidant enzyme activities in tumor cells. Prog. Med. Res. 3: 12.
- Przybyszewski W., Wideł M., Walichiewicz P., Snietura P., Lange D., Bkhiyan A. (2005): Protective effects of local temporary ischemia depends on applied dose of radiation. Cancer Lett. 222, 113-118.
- Ryabokon N., Goncharova R., Duburs G., Rzeszowska-Wolny J. (2005): 1,4-dihydropiridyne derivative reduces DNA damage and stimulates DNA repair in human cells in vitro. Mutation Res. (Gen. Tox. Environ. Mut.) 587: 52-58.
- Rzeszowska-Wolny J., Polanska J., Pietrowska M., Palyvoda O., Jaworska J., Butkiewicz D., Hancock R. (2005): Influence of Polymorphisms in DNA Repair Genes XPD, XRCC1 and MGMT on DNA Damage Induced by Gamma-Radiation and its Repair in Lymphocytes in vitro. Radiation Res., 164: 132-140.
- Ślosarek K., Konopacka M., Rogoliński J., Latocha M., Sochanik A. (2005): Effects of depth on radiation-induced cell damage in a water phantom Rep. Pract. Oncol. Radiother. 10: 37-41.
- Wideł M., Słowiński J., Mazurek U., Macyszyn G.B., Latocha M., Ligus J.G., Stomal M., Mrowka R. (2005): Cell proliferative activity by histone H2B mRNA level correlates with cytogenetic damage induced by radiation In human glioblastoma cell lines. Journal of Neuro-Oncology 71:237-243.
- Widłak P., Garrard W.T. (2005): Discovery, regulation, and action of the major apoptotic nucleases DFF40/CAD and endonuclease G. J. Cell. Biochem. 94, 1078-1087.
- Widłak P., Kalinowska M., Parseghian M.H., Lu X., Hansen J.C., Garrard W.T. (2005): The histone H1 C-terminal domain binds to the apoptotic nuclease, DNA Fragmentation Factor (DFF40/CAD) and stimulates DNA cleavage. Biochemistry 44, 7871-7878.
2004:
- Fedorov A., Lukyanov D., Rogoliński J., Widłak P., Podgorna O., Rzeszowska-Wolny J. (2004):The nuclear protein P30 specifically interacts with a nuclear matrix attachment region from the rat genome. Cell. Mol. Biol. Lett. 9, 153-165.
- Konopacka M. (2004):Rola witaminy C w uszkodzeniach oksydacyjych DNA. Postępy Hig. Med. Dośw. 58, 343-348.
- Konopacka M., Rogoliński J. (2004):Thiamine prevents X-ray induction of genetic changes in human lymphocytes in vitro. Acta Biochim. Polon. 51, 839-843.
- Łanuszewska J., Widłak P. (2004): The truncation of Ku86 in human lymphocytes. Cancer Letters 205, 197-205.
- Matulewicz Ł., Przybyszewski W.M. (2004): Wpływ lipidów na aktywność polimeraz DNA. Postępy Biologii Komórki 31, 277-284.
- Przybyszewski W.M., Wideł M., Szurko A., Lubecka B., Matulewicz Ł., Maniakowski Z., Polaniak R., Birkner E., Rzeszowska-Wolny J. (2004):Multiple bystander effect on irradiated megacolonies of melanoma cells on non-irradiated neighbours. Cancer Letters. 214, 91-102.
- Słowiński J., Bierzyńska-Macyszyn G., Mazurek U., Wideł M., Latocha M., Stomal M., Śnietura M., Mrówka R. (2004): Cytokinesis-block micronucleus assay in human glioma cells exposed to radiation. Image Anal. Stereol. 23, 159-165.
- Sokół M., Przybyszewski W.M., Matlas B. (2004):Investigation of metabolic changes in irradiated rat brain tissue by means of 1H NMR in vitro relaxation study. Solid State Nucl. Magn. Reson. 25, 53-60.
- Weil M.R., Widłak P., Minna J.D., Garner H.R. (2004): Global survey of chromatin accessibility using DNA microarrays. Genome Res.14, 1374-1381.
- Widłak P. (2004): DNA microarrays, a novel approach in studies of chromatin structure. Acta Biochim. Polon. 51, 1-8.
- Widłak W., Widłak P. (2004): MAR/SAR elements flank the rat HST70 gene transcription unit. Cell. Mol. Biol. Lett. 9, 123-133.
2003:
- Horak S., Polańska J., Widłak P. (2003):High levels of bulky DNA adducts in human sperm correlate with impaired fertility. Acta Bioch. Pol. 50, 197-203
- Horak S., Polańska J., Widłak P. (2003):Bulky DNA adducts in human sperm: relationship with fertility, semen quality, smoking, and environmental factors. Mutation Res. 537, 53-65.
- Konopacka M., Rzeszowska-Wolny J. (2003): Protective effects of vitamin C on radiation - induced DNA damage in cultured human lymphocytes. Human Monitoring for Genetic Effects. NATO Science Series I: Life and Behavioural Sciences, vol. 351, pp. 273-281.
- Kumala S., Niemiec P., Wideł M., Hancock R., Rzeszowska-Wolny J. (2003):Apoptosis and clonogenic survival in three tumour cell lines exposed to gamma rays or chemical genotoxic agents. Cell. Mol. Biol. Lett. 8, 515-525.
- Łanuszewska J., Widłak P. (2003):Białka detektorowe rozpoznające pęknięcia nici DNA i ich udział w mechanizmach komórkowej odpowiedzi na stres. Postępy Biochemii 49, 229-238.
- Palyvoda O., Polańska J., Wygoda A., Rzeszowska-Wolny J. (2003): DNA damage and repair in lymphocytes of normal indyviduals and cancer patients: studies by the comet assay and micronucleus tests. Acta Bioch. Pol. 50, 181-190.
- Świerniak A., Polański A., Śmieja J., Kimmel M., Rzeszowska-Wolny J. (2003): Asymptotic analysys of three random branching walk models arising in molecular biology. Control Cybernetics 32,
- Tarnawski R., Wideł M., Składowski K. (2003):Tumor cell repopulation during conventional and accelerated radiotherapy in the in vitro megacolony culture. Int J. Radiat. Oncol. Biol. Phys. J 55, 1074-1081
- Walichiewicz P., Przybyszewski W.M., Jochem J., Wideł M., Koterbicka A., Śnietura M. (2003):Inhibitory effect of local ischemic preconditioning in total body irradiated rats. Teratogen. Carcinogen. Mutagen. 23 (suppl.1), 195-205.
- Walichiewicz P., Przybyszewski W.M. (2003): Brachyterapia śródnaczyniowa tętnic obwodowych. Postępy Medycyny Klinicznej i Wojskowej 7, 20-24.
- Wideł M., Jędruś S., Łukaszczyk B., Raczek-Zwierzycka K., Świerniak A. (2003):Radiation - induced micronucleus frequency in peripheral blood lymphocytes is correlated with normal tissue damage in patients with cervical carcinoma undergoing radiotherapy. Radiat. Res. 159, 713-721.
- Widłak P., Łanuszewska J., Cary R.B., Garrard W.T. (2003):Subunit structures and stoichiometries of human DFF proteins before and after induction of apoptosis. J. Biol. Chem. 278, 26915-26922.
- Widłak P., Fujarewicz K. (2003): The analysis of chromatin condensation state and transcriptional activity using DNA microarrays. Journal of Medical Informatics and Technologies. 6, IP13-IP19.
2002:
- Beck E., Polaniak R., Wideł M., Drzazga Z. (2002): Influence of electromagnetic field on murine sqamous cell carcinoma cells in vitro. IFMBE Proceedings 2002, 2, 142-143.
- Konopacka M., Palyvoda O., Rzeszowska-Wolny J. (2002):Inhibitory effect of ascorbic acid post-treatment on radiation- induced chromosomal damage in human lymphocytes in vitro.Teratogen. Carcinogen. Mutagen. 22, 443-450.
- Konopacka M., Rzeszowska-Wolny J. (2002): "Bystander effect" - uszkodzenia genetyczne indukowane poza polem napromieniania. Nowotwory, 52. (suppl.3), 139-144.
- Palyvoda O., Mukalov I., Polańska J., Wygoda A., Drobot L., Wideł M., Rzeszowska-Wolny J. (2002): Radiation-induced DNA damage and its repair in lymphocytes of patients with head neck cancer and healthy donors. Anticancer Res. 22, 1721-1728.
- Przybyszewski W.M., Wideł M., Palyvoda O. (2002): Lipid peroxidation, DNA damage, and celluar morphology of R1 Rhabdomyosarcoma cell line irradiated in vitro by gamma-rays with different dose-rate. Teratogen. Carcinogen. Mutagen. 22, 93-102.
- Przybyszewski W.M., Walichiewicz P. (2002): Regulacja procesów wewnątrzkomórkowych przez kwasy tłuszczowe i końcowe produkty peroksydacji lipidów. Post. Hig. Med. Dośw. 56, 741-756.
- Przybyszewski W.M., Wideł M. (2002):Kwasy tłuszczowe jako potencjalny element wspomagający terapię nowotworów. Post. Hig. Med. Dośw. 56, 589-602.
- Walichiewicz P., Przybyszewski W.M. , Jochem J., Wideł M., Koterbicka A. (2002): Inhibitory effect of local ischemic preconditioning on gamma ray-induced lipid peroxidation in rats : a preliminary study. Int. J. Tissue React. 24, 143-150.
- Wideł M., Lubecka B., Tarnawski R., Czuba A. (2002): Fractionated radiotherapy of tumour megacolonies: Cytogenetic, flow cytometry and survival studies. IFMBE Proccedings 2002; 2, 190-191.
- Widłak P., Palyvoda O., Kumala S., Garrard W.T. (2002): Modeling apoptotic chromatin condensation in normal cell nuclei. J. Biol. Chem. 277, 21683-21690.
- Widłak P. (2002): Struktura chromatyny a powstawanie i naprawa uszkodzeń DNA. Kosmos, 51, 5-12.
2001:
- Fujarewicz K., Kimmel M., Rzeszowska-Wolny J., Świerniak A. (2001): Improved classification of microarray gene expression data using support vector machine. J. Med. Inform. Technol. 2, MI9-17.
- Konopacka M., Rzeszowska-Wolny J. (2001): Antioxidant Vitamins C, E and gamma-carotene reduce DNA damage before as well as after gamma-ray irradiation of human lymphocytes in vitro. Mutat. Res. 491, 1-7.
- Sokół M., Przybyszewski W.M., Pienążek P. Flakus L., Walecki J., Koterbicka A. (2001): Glucose detection - in vitro and in vivo 1H-NMR studies. Polish J. Med. Phys. Eng. 7, 111-117.
- Świerniak A., Kimmel M., Śmieja J., Rzeszowska-Wolny J. (2001): Modelling of cell aging - system theoretic approach. J. Med. Inform. Technol., 2, MI3-8.
- Wideł M., Kołosza Z., Jędruś S., Łukaszczyk B., Raczek-Zwierzycka K., Świerniak A. (2001):Micronucleus assay in vitro provides significant prognostic information in human cervical carcinoma; the updated analysis. Int. J. Radiat. Biol. 77, 631-635.
- Widłak P., Garrard W.T. (2001): Ionic and cofactor requirements for the activity of the apoptotic endonuclease DFF40/CAD. Mol. Cell. Biochem. 218, 125-130.
- Widłak P., Li L.Y., Wang X., Garrard W.T. (2001):Action of recombinant human apoptotic endonuclease G on naked DNA and chromatin substrates; cooperation with exonuclease and DNaseI. J. Biol. Chem., 276, 48404-48409.
2000:
- Chwałek A., Kramer G., Ratuszna A., Habdas J., Wideł M. (2000): Photodynamic effect of two water - souble porphyrins on human malignant melanoma and murine cervical carcinoma in multicellular megacolonies system in vitro. Polish J. Med. Phys. Eng. 6, 185-190.
- Fujarewicz K., Rzeszowska-Wolny J. (2000): Canser classification based on gene expresion data. J. Med. Inf. Tech. 5, BI23-BI27.
- Łanuszewska J., Widłak P. (2000): High mobility group 1 and 2 proteins bind preferentially to DNA that contains bulky adducts induced by benzo(a)pyrene diol epoxide and N-acetoxy-acetylaminofluorene. Canc. Lett. 158, 17-25.
- Łanuszewska J., Cudak A., Rzeszowska-Wolny J., Widłak P. (2000): Detection of damage - recognition proteins from human lymphocytes. Acta Bioch. Pol. 47, 443-450.
- Pietrowska M., Łanuszewska J., Walter Z., Rzeszowska-Wolny J., Widłak P. (2000): Detection and characterization of rat protein recognizing DNA damaged by N-acetoxy-acetylaminofluorene. Cell. Mol. Biol. Lett. 5, 423-431.
- Przybyszewski W.M. (2001): Udział produktów peroksydacji lipidów w przeciwnowotworowym mechanizmie działania promieniowania jonizującego i cytostatyków radiomimetycznych. Post. Hig. Med. Dośw. 55, 803-813.
- Rzeszowska-Wolny J. (2000): Sygnalizacja i zmiany funkcjonowania komórki po uszkodzeniu DNA. w: Na pograniczu chemii i biologii. Red. Korniak H., Barciszewski J. Tom IV, pp. 231-252.
- Widłak P., Li P., Wang X., Garrard W.T. (2000):Cleavage preferences of the apoptotic endonuclease DFF40 (Caspase - activated Dnase or Nuclease) on naked DNA and chromatin substrates. J. Biol. Chem. 275, 8226-8232.
- Widłak P. (2000): DFF40/CAD hypersensitive sites are potentially involved in high molecular weight DNA fragmentation during apoptosis. Cell. Mol. Biol. Lett. 5, 373-379.
- Widłak P. (2000): The DNA damage induced cell cycle checkpoints. J. Theor. Med. 2, 243-273.
- Widłak P. (2000): Naprawa dwunicowych pęknięć DNA indukowanych przez promieniowanie jonizujące. Post. Hig. Med. Dośw. 54, 133-148.
- Widłak P. (2000): Białka rozpoznające i wiążące się z uszkodzonym DNA; udział w mechanizmach naprawy DNA i kancerogenezie. Post. Biochem. 46, 198-206.
- Widłak P. (2000): Mechanizmy fragmentacji DNA i kondesacji chromatyny w komórkach ulegających apoptozie. Post. Biol. Kom. 27, 583-597.
- Widłak P. (2000): The DFF40/CAD endonuclease and its role in apoptosis. Acta Bioch. Pol. 47, 1037-1044.
1999:
- Liu X., Zou H., Widłak P., Garrard W.T, Wang X. (1999): Activation of the apoptotic endonuclease DFF40 (caspase-activated Dnase or nuclease). Oligomerization and direct interactions with histone H1. J. Biol. Chem. 274, 13836-13840.
- Łobos E., Polański A., Widłak P. (1999): On distribution of segregating sites in n-element samples of DNA sequences. Proceedings of the 5th National Conference on Application of Mathematics in Biology and Medicine. Ustrzyki, 1999, pp. 105-110.
- Pietrowska M., Rusin M., Widłak P., Razin S.V., Rzeszowska-Wolny J. (1999): Characterization of DNA sequences localized 11 to 17.5 kb upstream of the chicken embryonic pi-globin gene. Acta Biochim. Polon. 46, 774- 784.
- Rzeszowska-Wolny J., Widlak P. (1999):Damaged DNA-binding proteins: recognition of N-acetoxy-acetylaminofluorene-induced DNA adducts. Acta Bioch. Pol. 46, 173-180.
- Rzeszowska-Wolny J. (1999): Uszkodzenia DNA i ich rozpoznawanie w komórce. W: Na pograniczu chemii i biologii. Red. Korniak H., Barciszewski J. Tom III, pp. 113-139.
- Świerniak A., Rzeszowska-Wolny J., Kimmel M., Polański A. (1999): Asymptotic properties of microsatellite repeats model. Proceedings of the 5th National Conference on Application of Mathematics in Biology and Medicine. Ustrzyki. 1999, pp. 143-148.
- Świerniak A., Polański A., Kimmel M., Rzeszowska-Wolny J. (1999): Stability of infinite-dimensional model of microsatellite DNA repeats. IFAC Proc. 14th Triennal World Congress, Beijing, pp. 41-46.
- Świerniak A., Polański A., Rzeszowska-Wolny J. (1999): Control theoretic approach to gene amplification and microsatellite DNA repeats. Proceedings of 1999 Symp. on Robotics and Control, IEEE Hong-Kong, pp. 750- 755.
- Wideł M., Jędruś S., Owczarek S., Konopacka M., Lubecka B., Kołosza Z. (1999):The increment of micronucleus frequence in cervical carcinoma in vivo and its prognostic value for tumour radiocurability. Brit. J. Canc. 80, 1599-1608.
- Widłak P., Rzeszowska-Wolny J. (1999): Nuclear matrix and nucleotide excision repair: damage-recognition proteins are not constitutive components of the nuclear matrix. Gene Ther. Mol. Biol. 4, 275-284.
1998:
- Konopacka M., Wideł M., Rzeszowska-Wolny J. (1998):Modifing effect of vitamins C, E and beta-carotene against gamma-ray-induced DNA damage in mause cells. Mutation Res. 417, 85-94.
- Konopacka M., Wideł M., Rzeszowska-Wolny J. (1998): Antioxidant vitamins as modulators of DNA damage and repair. In: Progress in Radio-Oncology VI, eds. H.D. Kogelnik and F. Sedlmayer, Monduzzi Editore, pp. 233-236.
- Liu X., Li P., Widłak P., Zou H., Luo X., Garrard W.T., Wang X. (1998):DFF40 induces DNA fragmentation and chromatin condensation during apoptosis. Proc. Natl. Acad. Sci. USA 95, 8461-8466.
- Przybyszewski W.M., Wideł M., Koterbicka A. (1998): Peroxidative changes in rat heart myocardium produced by gamma rays or farmarubicin: prevention by cardioxane or neoton. Toxic Subst. Mech. 17, 1-14.
- Tarnawski R., Składowski K., Ficek K., Rzeszowska-Wolny J. (1998): Local control for head and neck cancer treated by radiation alone. Clinical radiobiology of hyperfractionation or accelerated radiotherapy. In: Progress in Radio-Oncology VI, eds. H.D. Kogelnik and F. Sedlmayer, Monduzzi Editore 1998, pp. 821-826.
- Wideł M., Przybyszewski M.W. (1998): Inverse Dose-Rate Effect for the Inducton of Micronuclei in Lewis Lung Carcinoma after Exposure to Cobalt-60 Gamma Rays. Radiation Res. 149, 98-102.
- Widłak P., Garrard W.T. (1998): Nucleosomes and regulation of gene expression. Structure of the HIV-1 5'LTR. Acta Bioch. Pol. 45, 209-219.
- Widłak P. (1998): Reconstitution of UV-damaged DNA into chromatin using Xenopus oocyte extracts. Acta Bioch. Pol. 45, 595-603.
- Widłak P. (1998): Apoptoza: nukleaza, kaspazy, cytochrom c. Post. Biochem. 44, 252-254.
1997:
- Przvbyszewski W.M., Wideł M., Koterbicka A. (1997): Failure of cardioxane (ICRF-187, Dexrazoxane) to limit peroxidative heart damage in rats after gamma irradiation or farmorubicin (4'-epidoxorubicin) treatment. Toxic Subst. Mech. 16, 133-149.
- Wideł M., Dobrut M., Mąka B., Lubecka B., Płuciennik A. (1997): The radiosensitivity of human malignant melanomas evaluated by cytokinesis-block micronucleus assay. Neoplasma 44, 109-116.
- Widłak P., Rogoliński J., Lisowska K., Rzeszowska-Wolny J. (1997): Sequences flanking rat HSP70 I gene contain motifs specific for matrix attached regions and form complexes with nuclear matrix protein in vitro. Cell. Mol. Biol. Let. 2, 317-324.
- Widłak P., Gaynor R.B., Garrard W.T. (1997):In vitro chromatin assembly of the HIV-1 promoter: ATP-dependent polar repositioning of nucleosomes by Sp1 and NFkappaB. J. Biol. Chem. 272, 17654-17661.
- Widłak P. (1997): Struktura nukleosomowa chromatyny a naprawa DNA. Post. Biol. Kom. 24, 325-337.
- Widłak P. (1997): Wpływ uszkodzeń DNA na regulację cyklu komórkowego. Post. Bioch. 43, 85-90.
Principal Investigators:
- Professor Joanna Rzeszowska, Ph.D., D.Sc.
Head of the Department
jwolny@io.gliwice.pl; jrzeszowskawolny@yahoo.com,
(+48 32) 278 96 77
- Assistant Professor Piotr Widłak, Ph.D., D.Sc.
widlak@io.gliwice.pl; pwidlak@yahoo.com,
(+48 32) 278 96 72
- Maria Wideł, Ph.D., D.Sc
widel@io.gliwice.pl,
(+48 32) 278 98 49
Research Personnel:
- Agnieszka Gdowicz, M.Sc.
agagdowicz@wp.pl, (+48 32) 278 96 27
- Robert Herok, M.Sc.
robertherok@yahoo.com, (+48 32) 278 96 26
- Magdalena Kalinowska, M.Sc.
mkalinowska@io.gliwice.pl, (+48 32) 278 96 74
- Maria Konopacka, Ph.D.
m_konopacka@pf.pl, (+48 32) 278 98 04
- Joanna Łanuszewska, Ph.D.
lanuszewska@wp.pl, (+48 32) 278 96 25
- Monika Pietrowska, Ph.D.
m_pietrowska@io.gliwice.pl, (+48 32) 278 96 25
- Waldemar Przybyszewski, Ph.D.
wmp@io.gliwice.pl, (+48 32) 278 98 12
- Jacek Rogoliński, Ph.D.
rogolinski@io.gliwice.pl, (+48 32) 278 98 04
- Katarzyna Szołtysek
szoltysek.k@wp.pl, (+48 32) 278 96 25
- Agnieszka Szurko, M.Sc.
agaspel@o2.pl, (+48 32) 278 96 32
Technical Personnel:
- Iwona Domińczyk
(+48 32) 278 96 25
- Dina Kobolowska - Secretary of the Department
(+48 32) 278 98 08
- Lucyna Ponge
(+48 32) 278 96 74
- Sylwia Drzewiecka
(+48 32) 278 96 74
Scholarship fellows:
- Nadzeya Ryabokon, Ph.D.
Institute of Genetics and Cytology National Academy of Sciences, Minsk, Belarus
(+48 32) 278 96 32
Students:
- Jakub Hanus
- Izabela Kołodziejczyk
- Karol Jelonek
- Krzysztof Pawełek
- Artur Cieśla - Pobuda
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