Home LiteratureArticle Details

PMID: 6209609 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Methylation of either cytosine in the recognition sequence CGCG inhibits ThaI cleavage of DNA.

Nucleic acids research ·Vol. 12 ·No. 21 ·1984-11-12 ·Pages 8073-83

Strobl JS, Thompson EB

Abstract

ThaI (CGCG) sites which overlap HhaI (GCGC) sites in phi X174 and pBR322 DNA were methylated in vitro with HhaI methylase and S-adenosylmethionine to yield CGmCG, mCGCG or mCGmCG (5-methylcytosine, mC). Methylation of either cytosine in the ThaI recognition sequence rendered the DNA resistant to ThaI cleavage. Rat pituitary cell genomic DNA was digested with ThaI or 2 other known methylation-sensitive enzymes, AvaI or XhoI. After electrophoresis and ethidium bromide straining of the DNA, all 3 enzymes showed the infrequent DNA cleavage characteristic of methylation-sensitive enzymes. Comparison of pituitary growth hormone (GH) genes bearing strain-specific degrees of methylation showed the less methylated gene to be more frequently cut by either AvaI or ThaI. ThaI resistant sites in GH genes were cleaved by ThaI after exposing cells to 5-azacytidine, an inhibitor of DNA methylation. We conclude that ThaI is a useful restriction enzyme for the analysis of mC at CGCG sequences in eukaryotic DNA.

MeSH Terms

5-Methylcytosine Adenoma Animals Azacitidine/pharmacology Base Sequence Cell Line Cytosine/analogs & derivatives DNA Restriction Enzymes/antagonists & inhibitors DNA, Neoplasm Deoxyribonucleases, Type II Site-Specific Methylation Oligodeoxyribonucleotides/pharmacology Oligonucleotides/pharmacology Pituitary Neoplasms Rats Substrate Specificity

Chemicals

DNA, Neoplasm Oligodeoxyribonucleotides Oligonucleotides 5-Methylcytosine Cytosine DNA Restriction Enzymes CGCG-specific type II deoxyribonucleases Deoxyribonucleases, Type II Site-Specific Azacitidine

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Strobl J S

Thompson E B

References (12)

12 references, click to expand

Nucleotide sequence of bacteriophage phi X174 DNA.
Nature. 1977 Feb 24;265(5596):687-95 PMID: 870828
A restriction enzyme Tha I from the thermophilic mycoplasma Thermoplasma acidophilum.
Nucleic Acids Res. 1978 Jun;5(6):1729-39 PMID: 673834
The integrated genome of murine leukemia virus.
Cell. 1978 Nov;15(3):1003-10 PMID: 728982
Nucleotide sequence specificity of restriction endonucleases.
Science. 1979 Aug 3;205(4405):455-62 PMID: 377492
Complete nucleotide sequence of the Escherichia coli plasmid pBR322.
Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:77-90 PMID: 383387
DNA methylation in the human gamma delta beta-globin locus in erythroid and nonerythroid tissues.
Cell. 1980 Apr;19(4):947-58 PMID: 6247075
Hormonal control of tyrosine aminotransferase, prolactin, and growth hormone induction in somatic cell hybrids.
J Steroid Biochem. 1980 Jan;12:193-210 PMID: 6106733
5-Methylcytosine in eukaryotic DNA.
Science. 1981 Jun 19;212(4501):1350-7 PMID: 6262918
The effect of site specific methylation on restriction endonuclease cleavage (update).
Nucleic Acids Res. 1983 Jan 11;11(1):r169-73 PMID: 6306558
Rat growth hormone expression in cell hybrids.
DNA. 1984;3(1):41-9 PMID: 6321121
On the mechanism of inhibition of DNA-cytosine methyltransferases by cytosine analogs.
Cell. 1983 May;33(1):9-10 PMID: 6205762
Establishment of clonal strains of rat pituitary tumor cells that secrete growth hormone.
Endocrinology. 1968 Feb;82(2):342-52 PMID: 4951281

Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/12.21.8073

Published

1984-11-12

Pages

8073-83

Language

English

Region

England

NLM ID

0411011

PMCID

PMC320278

Subset

External Links

PubMed source DOI: 10.1093/nar/12.21.8073 PMC Full Text View journal details

Analysis Services

Methylation of either cytosine in the recognition sequence CGCG inhibits ThaI cleavage of DNA.

Contact

WeChat Official Account