Methylation of deoxycytidine incorporated by excision-repair synthesis of DNA

Methylation of deoxycytidine incorporated by DNA excision-repair was studied in human diploid fibroblasts following damage with ultraviolet radiation, N-methyl-N-nitrosourea, or N-acetoxy-2-acetylaminofluorene. In confluent, nondividing cells, methylation in repair patches induced by all three agents is slow and incomplete. Whereas after DNA replication in logarithmic-phase cultures a steady state level of 3.4% 5-methylcytosine is reached in less than 2 hr after cells are labeled with 6- 3H-deoxycytidine, following ultraviolet-stimulated repair synthesis in confluent cells it takes about 3 days to reach a level of approximately 2.0% 5-methylcytosine in the repair patch. In cells from cultures in logarithmic-phase growth, 5-methylcytosine formation in ultraviolet-induced repair patches occurs faster and to a greater extent, reaching a level of approximately 2.7% in 10-20 hr. Preexisting hypomethylated repair patches in confluent cells are methylated further when the cells are stimulated to divide; however, the repair patch may still not be fully methylated before cell division occurs. Thus DNA damage and repair may lead to heritable loss of methylation at some sites.

[1]  E. Read,et al.  Methylation of DNA in HeLa cells after ultraviolet irradiation. , 1976, International journal of radiation oncology, biology, physics.

[2]  R. Burdon,et al.  The in vivo methylation of DNA in mouse fibroblasts. , 1969, Biochimica et biophysica acta.

[3]  R. Fuchs,et al.  Mechanism of inhibition of enzymatic deoxyribonucleic acid methylation by 2-(acetylamino)fluorene bound to deoxyribonucleic acid. , 1981, Biochemistry.

[4]  M. Ehrlich,et al.  Quantitative reversed-phase high performance liquid chromatographic determination of major and modified deoxyribonucleosides in DNA. , 1980, Nucleic acids research.

[5]  H. Cedar,et al.  Active gene sequences are undermethylated. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[6]  J. Kappler,et al.  The kinetics of DNA methylation in cultures of a mouse adrenal cell line , 1970, Journal of cellular physiology.

[7]  B. Puschendorf,et al.  Effect of hydroxyurea and trisethyleneiminobenzoquinone (Trenimon) on the methylation of DNA in Ehrlich ascites tumor cells. , 1972, Biochimica et biophysica acta.

[8]  R. Palmiter,et al.  DNA methylation controls the inducibility of the mouse metallothionein-I gene in lymphoid cells , 1981, Cell.

[9]  M. Groudine,et al.  Chromatin structure of endogenous retroviral genes and activation by an inhibitor of DNA methylation , 1981, Nature.

[10]  J. A. Elliott,et al.  Differences in removal of acetylaminofluorene and pyrimidine dimers from the DNA of cultured mammalian cells. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[11]  P. Cerutti,et al.  Excision of N-acetoxy-2-acetylaminofluorene-induced DNA adducts from chromatin fractions of human fibroblasts. , 1980, Cancer research.

[12]  T. Sneider,et al.  Repair methylation of parental DNA in synchronized cultures of Novikoff hepatoma cells. , 1975, Nucleic acids research.

[13]  H. Cedar,et al.  Organization of 5-methylcytosine in chromosomal DNA. , 1978, Biochemistry.

[14]  D E Graham,et al.  Analysis of repeating DNA sequences by reassociation. , 1974, Methods in enzymology.

[15]  T. Tlsty,et al.  Distribution of ultraviolet-induced DNA repair synthesis in nuclease sensitive and resistant regions of human chromatin. , 1978, Biochemistry.

[16]  R. Setlow,et al.  Rate and extent of DNA repair in nondividing human diploid fibroblasts. , 1981, Cancer research.

[17]  Michael R. Green,et al.  A unique pattern of integrated viral genes in hamster cells transformed by highly oncogenic human adenovirus 12 , 1976, Cell.

[18]  R. H. Stellwagen,et al.  5-Methylcytosine content of rat hepatoma DNA substituted with bromodeoxyuridine. , 1977, The Journal of biological chemistry.

[19]  A. Razin,et al.  DNA methylation and its possible biological roles. , 1981, Progress in nucleic acid research and molecular biology.

[20]  D. Billen,et al.  Methylation rapidly reannealing DNA during the cell cycle of Chinese hamster cells. , 1973, Biochimica et biophysica acta.

[21]  P. Price,et al.  Correlation between hypomethylation of DNA and expression of globin genes in Friend erythroleukemia cells. , 1977, European journal of biochemistry.

[22]  M. Lieberman,et al.  Deoxyribonucleoside incorporation during DNA repair of carcinogen-induced damage in human diploid fibroblasts. , 1973, Cancer research.

[23]  A. Bird,et al.  Use of restriction enzymes to study eukaryotic DNA methylation: II. The symmetry of methylated sites supports semi-conservative copying of the methylation pattern. , 1978, Journal of molecular biology.

[24]  D. Drahovsky,et al.  Hypomethylation of DNA in Raji cells after treatment with N-methyl-N-nitrosourea. , 1981, Carcinogenesis.

[25]  P. Hanawalt,et al.  Deficient repair of chemical adducts in α DNA of monkey cells , 1982, Cell.

[26]  I. Lacko,et al.  Enzymatic DNA methylation during repair synthesis in non-proliferating human peripheral lymphocytes. , 1976, Biochimica et biophysica acta.

[27]  M. Lieberman.  Approaches to the analysis of fidelity of DNA repair in mammalian cells. , 1976, International review of cytology.

[28]  A. Riggs,et al.  DNA methylation and gene function. , 1980, Science.

[29]  M. Lieberman,et al.  Distribution of deoxyribonucleic acid repair synthesis among repetitive and unique sequences in the human diploid genome. , 1974, Biochemistry.

[30]  W. Kreis,et al.  Inhibition of thymidylate synthase by hydroxyurea in rapidly proliferating P815 mastocytoma cells. , 1982, Biochimica et biophysica acta.

[31]  R. W. Davis,et al.  Rapid DNA isolations for enzymatic and hybridization analysis. , 1980, Methods in enzymology.

[32]  M. Kastan,et al.  Distribution of repair-incorporated nucleotides and nucleosome rearrangement in the chromatin of normal and xeroderma pigmentosum human fibroblasts. , 1979, Biochemistry.

[33]  T. Tlsty,et al.  The distribution of DNA repair synthesis in chromatin and its rearrangement following damage with N-acetoxy-2-acetylaminofluorene. , 1978, Nucleic acids research.