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PMID: 1187340 Published · ppublish English Journal Article

DNA methylase from HeLa cell nuclei.

Nucleic acids research ·Vol. 2 ·No. 10 ·1975-10-00 ·Pages 1669-84

Roy PH, Weissbach A

Abstract

A DNA methylase has been purified 270-fold from HeLa cell nuclei by chromatography on DEAE-cellulose, phosphocellulose, and hydroxyapatite. The enzyme transfers methyl groups from S-adenosyl-L-methionine to cytosine residues in DNA. The sole product of the reaction has been identified as 5-methylcytosine. The enzyme is able to methylate homologous (HeLa) DNA, although to a lesser extent than heterologous DNA. This may be due to incomplete methylation of HeLa DNA synthesized in vivo. The HeLa enzyme can methylate single-stranded DNA, and does so to an extent three times greater than that of the corresponding double-stranded DNA. In single-stranded M. luteus DNA, at least 2.4% of the cytosine residues can be methylated in vitro by the enzyme. The enzyme also can methylate poly (dG-dC-dG-dC) and poly (dG, dC). Bilateral nearest neighbors to the 5-methylcytosine have been determined with M. luteus DNA in vitro and HeLa DNA in vivo. The 5' neighbor can be either G or C while the 3' neighbor is always G and this sequence is, thus, p(G/C)pmCpG.

MeSH Terms
Cell Nucleus/analysis,enzymology DNA (Cytosine-5-)-Methyltransferases/isolation & purification,metabolism DNA, Neoplasm/analysis HeLa Cells/analysis,enzymology Kinetics Methyltransferases/metabolism Structure-Activity Relationship
Chemicals
DNA, Neoplasm Methyltransferases DNA (Cytosine-5-)-Methyltransferases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Roy P H
Weissbach A
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34 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/2.10.1669
Published
1975-10-00
Pages
1669-84
Language
English
Region
England
NLM ID
0411011
PMCID
PMC343536
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/2.10.1669 PMC Full Text View journal details
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