Human Placental DNA Methyltransferase: DNA Substrate and DNA Binding Specificity

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1984 Apr 25

PMID 6539466

Citations 12

Authors

R Y Wang

L H Huang

M Ehrlich

Affiliations

Soon will be listed here.

Abstract

We have partially purified a DNA methyltransferase from human placenta using a novel substrate for a highly sensitive assay of methylation of hemimethylated DNA. This substrate was prepared by extensive nick translation of bacteriophage XP12 DNA, which normally has virtually all of its cytosine residues replaced by 5-methylcytosine (m5C). Micrococcus luteus DNA was just as good a substrate if it was first similarly nick translated with m5dCTP instead of dCTP in the polymerization mixture. At different stages in purification and under various conditions (including in the presence or absence of high mobility group proteins), the methylation of m5C-deficient DNA and that of hemimethylated DNA were compared. Although hemimethylated , m5C-rich DNAs were much better substrates than were m5C-deficient DNAs and normal XP12 DNA could not be methylated, all of these DNAs were bound equally well by the enzyme. In contrast, from the same placental extract, a DNA-binding protein of unknown function was isolated which binds to m5C-rich DNA in preference to the analogous m5C-poor DNA.

Citing Articles

Mouse DNA-cytosine-5-methyltransferase: sequence specificity of the methylation reaction and electron microscopy of enzyme-DNA complexes.

Pfeifer G, Spiess E, Grunwald S, Boehm T, DRAHOVSKY D EMBO J. 1985; 4(11):2879-84.

PMID: 3933973 PMC: 554592. DOI: 10.1002/j.1460-2075.1985.tb04017.x.

Perturbation of maintenance and de novo DNA methylation in vitro by UVB (280-340 nm)-induced pyrimidine photodimers.

Becker F, Holton P, Ruchirawat M, Lapeyre J Proc Natl Acad Sci U S A. 1985; 82(18):6055-9.

PMID: 3862117 PMC: 390698. DOI: 10.1073/pnas.82.18.6055.

Kinetic mechanisms and interaction of rat liver DNA methyltransferase with defined DNA substrates.

Ruchirawat M, Noshari J, Lapeyre J Mol Cell Biochem. 1987; 76(1):45-54.

PMID: 3627114 DOI: 10.1007/BF00219397.

Effect of site-specific DNA methylation and mutagenesis on recognition by methylated DNA-binding protein from human placenta.

Zhang X, Ehrlich K, Wang R, Ehrlich M Nucleic Acids Res. 1986; 14(21):8387-97.

PMID: 3537958 PMC: 311866. DOI: 10.1093/nar/14.21.8387.

Transformation of Tetrahymena thermophila with hypermethylated rRNA genes.

Karrer K, Yao M Mol Cell Biol. 1988; 8(4):1664-9.

PMID: 3380092 PMC: 363326. DOI: 10.1128/mcb.8.4.1664-1669.1988.

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