DNA methyltransferases in normal and avian sarcoma virus-transformed rat cells. Quantitation of 5-methyldeoxycytidine in DNA and enzyme kinetics study.
In rat kidney cells transformed by avian sarcoma virus (B77 strain) DNA is hypomethylated (2.61 +/- 0.07%) when compared to DNA extracted from normal cells (3.33 +/- 0.11%) as revealed by high-performance liquid chromatography analysis. Kinetics studies showed that no significant differences could be detected between DNA methyltransferase activities from normal and transformed cells with regard to apparent Vmax, apparent Km for S-adenosylmethionine (2.32 X 10(-6) M and 6.64 X 10(-6) M respectively) and apparent Ki for S-adenosylhomocysteine (9.2 X 10(-7) M and 7.8 X 10(-7) M respectively), when unmethylated duplex DNA was used as second substrate. Equivalent ratios of S-adenosylmethionine over S-adenosylhomocysteine were measured in each cell type and DNA methyltransferase activities from both sources were found to be strictly additive. These results show that the hypomethylation of DNA detected in transformed cells is related neither to alterations of enzymatic activities extracted from nuclei nor to unbalanced S-adenosylmethionine/S-adenosylhomocysteine ratios.
D000818:Animals; D001358:Avian Sarcoma Viruses; D002467:Cell Nucleus / Q000201:enzymology; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000032:analysis*; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000096:biosynthesis; D005347:Fibroblasts / Q000201:enzymology*; D007668:Kidney; D007700:Kinetics; D051381:Rats; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism
D003841:Deoxycytidine; D012436:S-Adenosylmethionine; D004247:DNA; D012435:S-Adenosylhomocysteine; C016569:5-methyldeoxycytidine; D004248:DNA (Cytosine-5-)-Methyltransferases