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PMID: 8108416 Published · ppublish English Journal Article

Positioned nucleosomes inhibit Dam methylation in vivo.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 91 ·No. 4 ·1994-02-15 ·Pages 1361-5

Kladde MP, Simpson RT

Abstract

Escherichia coli Dam DNA methyltransferase can methylate genomic GATC sites when expressed in Saccharomyces cerevisiae. Others have observed changes in the level of methylation at specific sites and suggested that these changes are related to transcriptional state or chromosomal context. To test directly the influence of nucleosome location on the ability of Dam methyltransferase to modify GATC sites in chromatin, we analyzed minichromosomes containing precisely positioned nucleosomes in dam-expressing yeast strains. Levels of methylation at individual GATC sites were rigorously quantified by an oligonucleotide-probing procedure. Within the linker and adjacent 21 bp of nucleosome-associated DNA, GATC sites were highly methylated, whereas methylation was severely inhibited by histone-DNA contacts nearer to the nucleosomal pseudodyad. Other DNA-protein complexes also interfere with Dam methylation. These data are consistent with a model in which nucleosomes exert a repressive influence on the biological functions of DNA by restricting access of trans-acting factors to DNA.

MeSH Terms
Base Sequence DNA, Fungal/metabolism Deoxyribonucleases, Type II Site-Specific/metabolism Escherichia coli/enzymology Escherichia coli Proteins Methylation Methyltransferases/metabolism Models, Genetic Molecular Sequence Data Nucleosomes/physiology Recombinant Proteins/metabolism Saccharomyces cerevisiae/metabolism Site-Specific DNA-Methyltransferase (Adenine-Specific) Substrate Specificity Transcription, Genetic
Chemicals
DNA, Fungal Escherichia coli Proteins Nucleosomes Recombinant Proteins Methyltransferases Dam methyltransferase Site-Specific DNA-Methyltransferase (Adenine-Specific) dam protein, E coli endodeoxyribonuclease DpnI Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kladde M P
Laboratory of Cellular and Developmental Biology, National Institutes of Health, Bethesda, MD 20892.
Simpson R T
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.91.4.1361
Published
1994-02-15
Pages
1361-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC43158
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.91.4.1361 PMC Full Text View journal details
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