Publications
2004
Popiela A., Keith G., Borzecki A., Popiela G., Manowiec M., Gabrys M.
The meaning of the methylation of genomic DNA in the regulation of gene expression levels Article de journal
Dans: Eur J Gynaecol Oncol, vol. 25, no. 2, p. 145-9, 2004, (0392-2936 Journal Article Review Review, Tutorial).
Résumé | BibTeX | Étiquettes: *DNA, *Gene, Expression, Human, KEITH, Methylation, Neoplasms/*genetics, Neoplastic, Regulation
@article{,
title = {The meaning of the methylation of genomic DNA in the regulation of gene expression levels},
author = { A. Popiela and G. Keith and A. Borzecki and G. Popiela and M. Manowiec and M. Gabrys},
year = {2004},
date = {2004-01-01},
journal = {Eur J Gynaecol Oncol},
volume = {25},
number = {2},
pages = {145-9},
abstract = {INTRODUCTION: Methylation of genomic DNA is one of the major mechanisms that deactivates genes and regulates their tissue-specific transcription levels. Its patterns are based on clonal inheritance that occurs in the early stages of embryogenesis. All changes in the DNA methylation levels occurring especially in the promoter region of the genes, which involve hypo- as well as hyper-methylation, lead to cell differentiation and growth disorders. Therefore it can become an impulse that initiates different pathological processes including carcinogenesis. MATERIAL AND METHODS: The purpose of this review was to present the recent knowledge concerning methylation of genomic DNA based on recent references and authors' experience. RESULTS AND CONCLUSION: Genome stability disorders could be caused either by mutations, which damage the structure of the genes and have not been formerly removed, or as the consequence of an epigenetic mechanism. Methylation plays a decisive role in the activity of many genes and could be a natural weapon of an organism against the expression of foreign genetic material that degrades the original genome structure.},
note = {0392-2936
Journal Article
Review
Review, Tutorial},
keywords = {*DNA, *Gene, Expression, Human, KEITH, Methylation, Neoplasms/*genetics, Neoplastic, Regulation},
pubstate = {published},
tppubtype = {article}
}
Zukiel R., Nowak S., Barciszewska A. M., Gawronska I., Keith G., Barciszewska M. Z.
A simple epigenetic method for the diagnosis and classification of brain tumors Article de journal
Dans: Mol Cancer Res, vol. 2, no. 3, p. 196-202, 2004, (1541-7786 Journal Article).
Résumé | BibTeX | Étiquettes: *DNA, *Epigenesis, 5-Methylcytosine/*analysis, Adult, Aged, and, Brain, Chromatography, DNA, Female, Genetic, Gov't, Human, KEITH, Layer, Male, Methylation, Middle, Neoplasm/*chemistry/*metabolism, Neoplasms/*classification/*diagnosis/genetics/pathology, Non-U.S., Oxidative, oxygen, Reactive, Sensitivity, Species/metabolism, Specificity, Stress, Support, Thin
@article{,
title = {A simple epigenetic method for the diagnosis and classification of brain tumors},
author = { R. Zukiel and S. Nowak and A. M. Barciszewska and I. Gawronska and G. Keith and M. Z. Barciszewska},
year = {2004},
date = {2004-01-01},
journal = {Mol Cancer Res},
volume = {2},
number = {3},
pages = {196-202},
abstract = {The new, simple, and reliable method for the diagnosis of brain tumors is described. It is based on a TLC quantitative determination of 5-methylcytosine (m(5)C) in relation to its damage products of DNA from tumor tissue. Currently, there is evidence that oxidative stress through reactive oxygen species (ROS) plays an important role in the etiology and progression of several human diseases. Oxidative damage of DNA, lipids, and proteins is deleterious for the cell. m(5)C, along with other basic components of DNA, is the target for ROS, which results in the appearance of new modified nucleic acid bases. If so, m(5)C residue constitutes a mutational hotspot position, whether it occurs within a nucleotide sequence of a structural gene or a regulatory region. Here, we show the results of the analysis of 82 DNA samples taken from brain tumor tissues. DNA was isolated and hydrolyzed into nucleotides, which, after labeling with [gamma-(32)P]ATP, were separated on TLC. Chromatograms were evaluated using PhosphorImager and the amounts of 5-methyldeoxycytosine (m(5)dC) were calculated as a ratio (R) of m(5)dC to m(5)dC + deoxycytosine + deoxythymidine spot intensities. The R value could not only be a good diagnostic marker for brain tumors but also a factor differentiating low-grade and high-grade gliomas. Therefore, DNA methylation pattern might be a useful tool to give a primary diagnosis of a brain tumor or as a marker for the early detection of the relapse of the disease. This method has several advantages over those existing nowadays.},
note = {1541-7786
Journal Article},
keywords = {*DNA, *Epigenesis, 5-Methylcytosine/*analysis, Adult, Aged, and, Brain, Chromatography, DNA, Female, Genetic, Gov't, Human, KEITH, Layer, Male, Methylation, Middle, Neoplasm/*chemistry/*metabolism, Neoplasms/*classification/*diagnosis/genetics/pathology, Non-U.S., Oxidative, oxygen, Reactive, Sensitivity, Species/metabolism, Specificity, Stress, Support, Thin},
pubstate = {published},
tppubtype = {article}
}
2003
Miturski R., Postawski K., Semczuk A., Bogusiewicz M., Baranowski W., Jakowicki J. A., Keith G.
Global DNA methylation in relation to hMLH1 and hMSH2 protein immunoreactivity in sporadic human endometrial carcinomas Article de journal
Dans: Int J Mol Med, vol. 11, no. 5, p. 569-74, 2003, (1107-3756 Journal Article).
Résumé | BibTeX | Étiquettes: *DNA, Base, Carcinoma/genetics/*metabolism/pathology, DNA, Endometrial, Female, Gov't, Human, Immunohistochemistry, Methylation, Mismatch, Neoplasm, Neoplasms/genetics/*metabolism/pathology, Non-U.S., Pair, Proteins/*metabolism, Proto-Oncogene, Repair, Support
@article{,
title = {Global DNA methylation in relation to hMLH1 and hMSH2 protein immunoreactivity in sporadic human endometrial carcinomas},
author = { R. Miturski and K. Postawski and A. Semczuk and M. Bogusiewicz and W. Baranowski and J. A. Jakowicki and G. Keith},
year = {2003},
date = {2003-01-01},
journal = {Int J Mol Med},
volume = {11},
number = {5},
pages = {569-74},
abstract = {Overall DNA methylation status was studied in a group of 28 sporadic human endometrial carcinomas (ECs) using the [32P]-postlabeling technique. Moreover, expression of the DNA mismatch repair proteins (hMLH1 and hMSH2) was investigated in ECs using immunohistochemistry. Mean 5-methyldeoxycytosine (m5dC) content in the studied group was 3.48+/-0.37% (range, 2.89-4.12%). The mean m5dC scores were significantly different between early (3.35+/-0.33%) and advanced (3.66+/-0.36%) endometrial neoplasms (chi2-test; p=0.03). There was a markedly increased overall DNA methylation with the degree of histological differentiation and with the infiltration of the myometrium (p<0.05). Loss of hMLH1 and hMSH2 expression was reported in 7 (25%) and 5 (18%) tumors, respectively, but the immunoreactivity did not correlate with the known clinicopathological variables of cancer. In addition, no obvious correlation was found between global m5dC content and the lack of hMLH1 and hMSH2 protein expression in human uterine tumors (p=0.97 and p=0.19 for hMLH1 and hMSH2, respectively; Spearman's rank correlation test). Our results clearly show that alterations in global DNA methylation may influence tumor progression, but they are not directly associated with the inactivation of the mismatch-repair machinery in sporadic human ECs.},
note = {1107-3756
Journal Article},
keywords = {*DNA, Base, Carcinoma/genetics/*metabolism/pathology, DNA, Endometrial, Female, Gov't, Human, Immunohistochemistry, Methylation, Mismatch, Neoplasm, Neoplasms/genetics/*metabolism/pathology, Non-U.S., Pair, Proteins/*metabolism, Proto-Oncogene, Repair, Support},
pubstate = {published},
tppubtype = {article}
}
2002
Popiela A., Gabrys M. S., Rabczynski J., Panszczyk M., Keith G., Baranowski W.
[Estimation of DNA methylation level in endometrial cancer tissues] Article de journal
Dans: Ginekol Pol, vol. 73, no. 11, p. 966-9, 2002, (0017-0011 Journal Article).
Résumé | BibTeX | Étiquettes: *DNA, 80, Abstract, Adenocarcinoma/chemistry/*genetics, Aged, and, Case-Control, Endometrial, English, Expression, Female, Gene, Human, Hyperplasia/genetics, Methylation, Middle, Neoplasms/chemistry/*genetics, Neoplastic, over, Regulation, Studies
@article{,
title = {[Estimation of DNA methylation level in endometrial cancer tissues]},
author = { A. Popiela and M. S. Gabrys and J. Rabczynski and M. Panszczyk and G. Keith and W. Baranowski},
year = {2002},
date = {2002-01-01},
journal = {Ginekol Pol},
volume = {73},
number = {11},
pages = {966-9},
abstract = {OBJECTIVES: A level of DNA methylation plays an important role in regulation of cellular gene's expression. Estimation of DNA methylation level in endometrial neoplastic tissues compared to normal endometrial samples was the aim of this study. DESIGN: It was to be shown, that changes in methylation rate in promotory regions could lead to carcinogenesis in particular cell. Authors describe an analysis of DNA methylation level in endometrial cancer tissues compared to DNA methylation level in normal or hyperplastic endometrium. MATERIAL AND METHODS: Endometrial samples from 88 women were collected. 56 of them were classified as adenocarcinoma, 20 as hyperplastic changes, 12 as normal endometrium-control group. DNA was isolated from tissues and than prepared to pm5dC and pdC. Than we performed a statistical analysis of results. RESULTS: The median DNA methylation level was significantly higher in neoplastic tissues than in normal endometrium. There was no difference between DNA methylation level between normal endometrium and hyperplastic changes. CONCLUSIONS: Authors conclude that neoplastic endometrial tissues show high DNA methylation rate compared to normal or hyperplastic endometrium.},
note = {0017-0011
Journal Article},
keywords = {*DNA, 80, Abstract, Adenocarcinoma/chemistry/*genetics, Aged, and, Case-Control, Endometrial, English, Expression, Female, Gene, Human, Hyperplasia/genetics, Methylation, Middle, Neoplasms/chemistry/*genetics, Neoplastic, over, Regulation, Studies},
pubstate = {published},
tppubtype = {article}
}
Popiela A., Gabrys M. S., Rabczynski J., Panszczyk M., Keith G., Baranowski W.
[Estimation of DNA methylation level in nonendometrial uterus malignancies] Article de journal
Dans: Ginekol Pol, vol. 73, no. 11, p. 962-5, 2002, (0017-0011 Journal Article).
Résumé | BibTeX | Étiquettes: *DNA, 80, Abstract, Age, Aged, and, Case-Control, English, Expression, Factors, Female, Gene, Human, Mesodermal/genetics/*metabolism, Methylation, Middle, Mixed, Neoplasms/genetics/*metabolism, Neoplastic, over, Regulation, silencing, Studies, tumor, Uterine
@article{,
title = {[Estimation of DNA methylation level in nonendometrial uterus malignancies]},
author = { A. Popiela and M. S. Gabrys and J. Rabczynski and M. Panszczyk and G. Keith and W. Baranowski},
year = {2002},
date = {2002-01-01},
journal = {Ginekol Pol},
volume = {73},
number = {11},
pages = {962-5},
abstract = {OBJECTIVES: Rebuilding of genome structure leads to many pathological states including neoplastic malignancies. Rebuilding often occurs as a process caused by disturbances in gene silencing mechanism. DNA methylation pattern is one of the most important mechanisms connected to gene's silencing. Estimation of DNA methylation level in nonendometrial uterine neoplastic tissues compared to normal endometrial samples was the aim of this study. DESIGN: It was to be shown, that changes in methylation rate in promotory regions could lead to carcinogenesis in particular cell. Authors describe an analysis of DNA methylation level in nonendometrial neoplastic uterine tissues compared to DNA methylation level in normal endometrium. MATERIALS AND METHODS: Tissue samples from 9 women with tumor mixtus mesodermalis were collected. 12 samples were normal endometrium-control group. DNA was isolated from tissues and than we performed an estimation of DNA methylation levels. Than we performed a statistical analysis of results. RESULTS: The median DNA methylation level was significantly higher in neoplastic tissues than in normal endometrium. CONCLUSIONS: Authors conclude, that DNA methylation level is higher in neoplastic tissues, but does not correlate with clinical stage of the disease.},
note = {0017-0011
Journal Article},
keywords = {*DNA, 80, Abstract, Age, Aged, and, Case-Control, English, Expression, Factors, Female, Gene, Human, Mesodermal/genetics/*metabolism, Methylation, Middle, Mixed, Neoplasms/genetics/*metabolism, Neoplastic, over, Regulation, silencing, Studies, tumor, Uterine},
pubstate = {published},
tppubtype = {article}
}
1999
Wilhelm M., Heyman T., Boutabout M., Wilhelm F. X.
A sequence immediately upstream of the plus-strand primer is essential for plus-strand DNA synthesis of the Saccharomyces cerevisiae Ty1 retrotransposon Article de journal
Dans: Nucleic Acids Res, vol. 27, no. 23, p. 4547-52, 1999, (0305-1048 Journal Article).
Résumé | BibTeX | Étiquettes: *DNA, *Retroelements, Base, cerevisiae/*genetics, DNA, Elements, Fungal/*biosynthesis, Gov't, Mutation, Non-U.S., Primers, response, Saccharomyces, Sequence, Support
@article{,
title = {A sequence immediately upstream of the plus-strand primer is essential for plus-strand DNA synthesis of the Saccharomyces cerevisiae Ty1 retrotransposon},
author = { M. Wilhelm and T. Heyman and M. Boutabout and F. X. Wilhelm},
year = {1999},
date = {1999-01-01},
journal = {Nucleic Acids Res},
volume = {27},
number = {23},
pages = {4547-52},
abstract = {Priming of plus-strand DNA is a critical step in reverse transcription of retroviruses and retrotransposons. All retroelements use an RNase H-resistant oligoribonucleotide spanning a purine-rich sequence (the polypurine tract or PPT) to prime plus-strand DNA synthesis. Plus-strand DNA synthesis of the yeast Saccharomyces cerevisiae Ty1-H3 retrotransposon is initiated at two sites, PPT1 and PPT2, located at the upstream boundary of the 3'-long terminal repeat and near the middle of the pol gene in the integrase coding region. The two plus-strand primers have the same purine-rich sequence GGGTGGTA. This sequence is not sufficient by itself to generate a plus-strand origin since two identical sequences located upstream of PPT2 in the integrase coding region are not used efficiently as primers for plus-strand DNA synthesis. Thus, other factors must be involved in the formation of a specific plus-strand DNA primer. We show here that mutations upstream of the PPT in a highly conserved T-rich region severely alters plus-strand DNA priming of Ty1. Our results demonstrate the importance of sequences or structural elements upstream of the PPT for initiation of plus-strand DNA synthesis.},
note = {0305-1048
Journal Article},
keywords = {*DNA, *Retroelements, Base, cerevisiae/*genetics, DNA, Elements, Fungal/*biosynthesis, Gov't, Mutation, Non-U.S., Primers, response, Saccharomyces, Sequence, Support},
pubstate = {published},
tppubtype = {article}
}
1998
Duranton B., Keith G., Goss, Bergmann C., Schleiffer R., Raul F.
Concomitant changes in polyamine pools and DNA methylation during growth inhibition of human colonic cancer cells Article de journal
Dans: Exp Cell Res, vol. 243, no. 2, p. 319-25, 1998, (0014-4827 Journal Article).
Résumé | BibTeX | Étiquettes: *Cell, *DNA, &, Adenosylmethionine, Amidines/pharmacology, Caco-2, Cells, Decarboxylase/antagonists, Differentiation/drug, Division/drug, effects, Eflornithine/pharmacology, enzyme, Gov't, Human, Indans/pharmacology, inhibitors/metabolism, Inhibitors/pharmacology, Methylation, Non-U.S., Ornithine, Polyamines/*metabolism, Support
@article{,
title = {Concomitant changes in polyamine pools and DNA methylation during growth inhibition of human colonic cancer cells},
author = { B. Duranton and G. Keith and Goss and C. Bergmann and R. Schleiffer and F. Raul},
year = {1998},
date = {1998-01-01},
journal = {Exp Cell Res},
volume = {243},
number = {2},
pages = {319-25},
abstract = {The effects of CGP 48664 and DFMO, selective inhibitors of the key enzymes of polyamine biosynthesis, namely, of S-adenosylmethionine decarboxylase (AdoMetDC) and ornithine decarboxylase (ODC), were investigated on growth, polyamine metabolism, and DNA methylation in the Caco-2 cell line. Both inhibitors caused growth inhibition and affected similarly the initial expression of the differentiation marker sucrase. In the presence of the AdoMetDC inhibitor, ODC activity and the intracellular pool of putrescine were enhanced, whereas the spermidine and spermine pools were decreased. In the presence of the ODC inhibitor, the AdoMetDC activity was enhanced and the intracellular pools of putrescine and spermidine were decreased. With both compounds, the degree of global DNA methylation was increased. Spermine and spermidine (but not putrescine) selectively inhibited cytosine-DNA methyltransferase activity. Our observations suggest that spermidine (and to a lesser extent spermine) controls DNA methylation and may represent a crucial step in the regulation of Caco-2 cell growth and differentiation.},
note = {0014-4827
Journal Article},
keywords = {*Cell, *DNA, &, Adenosylmethionine, Amidines/pharmacology, Caco-2, Cells, Decarboxylase/antagonists, Differentiation/drug, Division/drug, effects, Eflornithine/pharmacology, enzyme, Gov't, Human, Indans/pharmacology, inhibitors/metabolism, Inhibitors/pharmacology, Methylation, Non-U.S., Ornithine, Polyamines/*metabolism, Support},
pubstate = {published},
tppubtype = {article}
}
1996
Friant S., Heyman T., Wilhelm F. X., Wilhelm M.
Role of RNA primers in initiation of minus-strand and plus-strand DNA synthesis of the yeast retrotransposon Ty1 Article de journal
Dans: Biochimie, vol. 78, no. 7, p. 674-80, 1996, (0300-9084 Journal Article).
Résumé | BibTeX | Étiquettes: *DNA, Acid, Bacterial/*metabolism, Complementary/metabolism, Conformation, Data, DNA, Elements, Gov't, Molecular, Mutagenesis, Non-U.S., Nucleic, Replication, RNA, RNA/*metabolism, Sequence, Support, Transposable
@article{,
title = {Role of RNA primers in initiation of minus-strand and plus-strand DNA synthesis of the yeast retrotransposon Ty1},
author = { S. Friant and T. Heyman and F. X. Wilhelm and M. Wilhelm},
year = {1996},
date = {1996-01-01},
journal = {Biochimie},
volume = {78},
number = {7},
pages = {674-80},
abstract = {The Ty1 retrotransposon of the yeast Saccharomyces cerevisiae is a long terminal repeat mobile genetic element that transposes through an RNA intermediate. Initiation of minus-strand and plus-strand DNA synthesis are two critical steps during reverse transcription of the retrotransposon genome. Initiation of minus-strand DNA synthesis of the Ty1 element is primed by the cytoplasmic initiator methionine tRNA base paired to the primer binding site near the 5' end of the genomic RNA. A structural probing study of the primer tRNA-Ty1 RNA binary complex reveals that besides interactions between the primer binding site and the last 10 nucleotides at the 3' end of the primer tRNA, three short regions of Ty1 RNA named box 0, box 1 and box 2.1 interact with the T and D stems and loops of the primer tRNA. Some in vivo results underline the functional importance of the nucleotide sequence of the boxes and suggest that extended interactions between genomic Ty1 RNA and the primer tRNA play a role in the reverse transcription pathway. Plus-strand DNA synthesis is initiated from an RNase H resistant oligoribonucleotide spanning a purine-rich sequence, the polypurine tract (PPT). Two sites of initiation located at the 5' boundary of the 3' long terminal repeat (PPT1) and near the middle of the TyB (pol) gene in the integrase coding sequence (PPT2) have been identified in the genome of Ty1. The two PPTs have an identical sequence, TGGGTGGTA. Mutations replacing purines by pyrimidines in this sequence significantly diminish or abolish initiation of plus-strand DNA synthesis. Ty1 elements bearing a mutated PPT2 sequence are not defective for transposition whereas mutations in PPT1 abolish transposition.},
note = {0300-9084
Journal Article},
keywords = {*DNA, Acid, Bacterial/*metabolism, Complementary/metabolism, Conformation, Data, DNA, Elements, Gov't, Molecular, Mutagenesis, Non-U.S., Nucleic, Replication, RNA, RNA/*metabolism, Sequence, Support, Transposable},
pubstate = {published},
tppubtype = {article}
}
1995
Heyman T., Agoutin B., Friant S., Wilhelm F. X., Wilhelm M. L.
Plus-strand DNA synthesis of the yeast retrotransposon Ty1 is initiated at two sites, PPT1 next to the 3' LTR and PPT2 within the pol gene. PPT1 is sufficient for Ty1 transposition Article de journal
Dans: J Mol Biol, vol. 253, no. 2, p. 291-303, 1995, (0022-2836 Journal Article).
Résumé | BibTeX | Étiquettes: *DNA, *Genes, *Repetitive, *Retroelements, Acid, Base, C/analysis, cerevisiae/genetics/*virology, Chain, Cloning, Data, DNA, Fungal, Fungal/biosynthesis, Genes, Genetic, Genome, Gov't, Mapping, Molecular, Non-U.S., Nucleic, pol, Poly, Polymerase, Primers, Reaction, Replication, Restriction, Saccharomyces, Sequence, Sequences, Support, Transcription, Viral, Viral/*biosynthesis
@article{,
title = {Plus-strand DNA synthesis of the yeast retrotransposon Ty1 is initiated at two sites, PPT1 next to the 3' LTR and PPT2 within the pol gene. PPT1 is sufficient for Ty1 transposition},
author = { T. Heyman and B. Agoutin and S. Friant and F. X. Wilhelm and M. L. Wilhelm},
year = {1995},
date = {1995-01-01},
journal = {J Mol Biol},
volume = {253},
number = {2},
pages = {291-303},
abstract = {Long terminal repeat elements and retroviruses require primers for initiation of minus and plus-strand DNA synthesis by reverse transcriptase. Here we demonstrate genetically that plus-strand DNA synthesis of the yeast Ty1 element is initiated at two sites located at the 5' boundary of the 3' long terminal repeat (PPT1) and near the middle of the pol gene in the integrase coding sequence (PPT2). A consequence of the presence of two PPTs is that Ty1 plus-strand DNA exists as segments at some time during replication. Three fragments have been identified: the plus-strand strong-stop DNA initiated at PPT1, a downstream fragment initiated at PPT2 and an upstream fragment spanning the 5'-terminal part of Ty1 and a portion of the TyB gene. Characterization of the 3' ends of the plus-strand DNA fragments reveals (1) that the upstream fragment is elongated beyond PPT2 creating a plus-strand overlap and (2) that the majority of plus-strand strong-stop DNA fragments bear a copy of the minus-strand primer binding site in agreement with the accepted model of retroviral genomic RNA reverse transcription. The two polypurine tracts, PPT1 and PPT2, have an identical sequence GGGTGGTA. Mutations replacing purines by pyrimidines in this sequence significantly diminish or abolish initiation of plus-strand synthesis. Ty1 elements bearing a mutated PPT2 sequence are not defective for transposition whereas mutations in PPT1 abolish transposition.},
note = {0022-2836
Journal Article},
keywords = {*DNA, *Genes, *Repetitive, *Retroelements, Acid, Base, C/analysis, cerevisiae/genetics/*virology, Chain, Cloning, Data, DNA, Fungal, Fungal/biosynthesis, Genes, Genetic, Genome, Gov't, Mapping, Molecular, Non-U.S., Nucleic, pol, Poly, Polymerase, Primers, Reaction, Replication, Restriction, Saccharomyces, Sequence, Sequences, Support, Transcription, Viral, Viral/*biosynthesis},
pubstate = {published},
tppubtype = {article}
}