Human DNA-demethylating activity: a glycosylase associated with RNA and PCNA.
We have partially purified and characterized the 5-methylcytosine removing activity (5-meC-DNA Glycosylase) from HeLa cells with 700-fold enrichment. This activity cleaves DNA specifically at fully methylated CpG sites. The mechanism of 5-meC removal is base excision from fully methylated CpG loci on DNA, producing abasic sites. Hemi-methylated DNA is not a substrate. A prominent 52 KDa protein is present in all partially purified fractions. This activity is tightly associated with other nuclear factors and proteins, which resulted in differential fractionation of this activity on ion exchange columns. One nuclear factor associated with this activity is identified as RNA. Another nuclear protein, proliferating cell nuclear antigen (PCNA) is also associated with this enzyme. Glycosylic removal of 5-meC from DNA by this activity could be involved in the regulation of transcription, replication, differentiation, and development through resultant hypomethylation of DNA.
Vairapandi
M
M
The Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140, USA. vairam@unix.temple.edu
Liebermann
D A
DA
Hoffman
B
B
Duker
N J
NJ
D001483:Base Sequence; D018899:CpG Islands; D045647:DNA Glycosylases; D019175:DNA Methylation; D004277:DNA, Single-Stranded; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D013379:Substrate Specificity
D004277:DNA, Single-Stranded; D018809:Proliferating Cell Nuclear Antigen; D012313:RNA; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases