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Evidence for gene silencing by endogenous DNA methylation

By Robin Holliday and Thu Ho

Abstract

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-[(3)H]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(+)

Topics: Biological Sciences
Publisher: The National Academy of Sciences
Year: 1998
DOI identifier: 10.1073/pnas.95.15.8727
OAI identifier: oai:pubmedcentral.nih.gov:21144
Provided by: PubMed Central
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