Evidence for gene silencing by endogenous DNA methylation.
Transformed cells can spontaneously silence genes by de novo methylation, and it is generally assumed that this is due to DNA methyltransferase activity. We have tested the alternative hypothesis that gene silencing could be due to the uptake of 5-methyl-dCMP into DNA, via the di- and triphosphonucleotides. 5-Methyl-dCMP would be present in cells from the ongoing repair of DNA. We have isolated a strain of Chinese hamster ovary (CHO) cells, designated HAM-, which spontaneously silences two tested genes at a very high frequency. We have shown that this strain incorporates 5-[3H]methyldeoxycytidine into 5-methylcytosine and thymine in DNA. It also has low 5-methyl-dCMP deaminase activity. Another HAM+ strain has high deaminase activity and a very low frequency of gene silencing. The starting strain, CHO K1, has a phenotype intermediate between HAM- and HAM+.
Commonwealth Scientific and Industrial Research Organization, Molecular Science, Sydney Laboratory, P.O. Box 184, North Ryde NSW 2113, Australia.
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D000818:Animals; D016466:CHO Cells; D002455:Cell Division; D006224:Cricetinae; D003470:Culture Media; D004247:DNA / Q000378:metabolism; D019175:DNA Methylation; D003843:Deoxycytidine Monophosphate / Q000031:analogs & derivatives / Q000378:metabolism; D005816:Genetic Complementation Test
C113700:5-methyldeoxycytidine monophosphate; D003470:Culture Media; D003843:Deoxycytidine Monophosphate; D004247:DNA