Detection of 5-methylcytosine in DNA sequences.

Nucleic acids research ·Vol. 5 ·No. 5 ·1978-05-00 ·Pages 1479-85

Ohmori H, Tomizawa JI, Maxam AM

Abstract

Col E1 DNA has methylated cytosine in the sequence 5'-CC*(A/T)GG-3' and methylated adenine in the sequence 5'-GA*TC-3' at the positions indicated by asterisks(*). When the Maxam-Gilbert DNA sequencing method is applied to this DNA, the methylated cytosine (5-methylcytosine) is found to be less reactive to hydrazine than are cytosine and thymine, so that a band corresponding to that base does not appear in the pyrimidine cleavage patterns. The existence of the methylated cytosine can be confirmed by analyzing the complementary strand or unmethylated DNA. In contrast, the methylated adenine (probably N6-methyladenine) cannot be distinguished from adenine with standard conditions for cleavage at adenine.

MeSH Terms

Base Sequence Cytosine/analogs & derivatives,analysis DNA Restriction Enzymes DNA, Bacterial Escherichia coli/analysis

Chemicals

DNA, Bacterial Cytosine DNA Restriction Enzymes

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Ohmori H

Tomizawa J I

Maxam A M

References (16)

16 references, click to expand

STUDIES OF THE NUCLEOTIDE ARRANGEMENT IN DEOXYRIBONUCLEIC ACIDS. IX. SELECTIVE DEGRADATION OF PYRIMIDINE DEOXYRIBONUCLEOTIDES.
Biochim Biophys Acta. 1964 Nov 15;91:462-76 PMID: 14254018
In vivo methylation by Escherichia coli K-12 mec+ deoxyribonucleic acid-cytosine methylase protects against in vitro cleavage by the RII restriction endonuclease (R. Eco RII).
J Bacteriol. 1976 May;126(2):990-6 PMID: 770462
Complementary specificity of restriction endonucleases of Diplococcus pneumoniae with respect to DNA methylation.
J Mol Biol. 1977 Jul;114(1):153-68 PMID: 20509
Studies on the biological role of DNA methylation. II. Role of phiX174 DNA methylation in the process of viral progeny DNA synthesis.
Nucleic Acids Res. 1976 Oct;3(10):2665-75 PMID: 136644
Origin of replication of colicin E1 plasmid DNA.
Proc Natl Acad Sci U S A. 1977 May;74(5):1865-9 PMID: 325558
A new method for sequencing DNA.
Proc Natl Acad Sci U S A. 1977 Feb;74(2):560-4 PMID: 265521
Two sequence-specific endonucleases from Moraxella bovis.
J Mol Biol. 1977 Jul;114(1):169-79 PMID: 909085
DNA substrate site for the EcoRII restriction endonuclease and modification methylase.
Nat New Biol. 1973 Jul 11;244(132):40-3 PMID: 4124385
Recognition sequence of a restriction enzyme.
Nat New Biol. 1973 Jul 4;244(131):7-10 PMID: 4578426
Location of DNA methylation genes on the Escherichia coli K-12 genetic map.
Mol Gen Genet. 1973 Dec 14;127(1):47-55 PMID: 4589344
Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form.
Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159-66 PMID: 4894690
The degradation of DNA by hydrazine: a critical study of the suitability of the reaction for the quantitative determination of purine nucleotide sequences.
Biochim Biophys Acta. 1969 Feb 18;174(2):591-603 PMID: 4888872
Deoxyribonucleic acid-cytosine methylation by host- and plasmid-controlled enzymes.
J Bacteriol. 1975 Apr;122(1):129-38 PMID: 1091619
A specific endonuclease from Haemophilus haemolyticus.
J Mol Biol. 1976 May 5;103(1):199-208 PMID: 1085372
The sensitivity of bacteriophage lambda DNA to restriction endonuclease RII.
J Mol Biol. 1975 Nov 5;98(3):645-7 PMID: 1104876
A specific partial hydrolysis procedure for soluble RNA.
Biochim Biophys Acta. 1963 Mar 26;68:484-6 PMID: 13996927

Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/5.5.1479

Published

1978-05-00

Pages

1479-85

Language

English

Region

England

NLM ID

0411011

PMCID

PMC342097

Subset

External Links

PubMed source DOI: 10.1093/nar/5.5.1479 PMC Full Text View journal details

Analysis Services

Detection of 5-methylcytosine in DNA sequences.

Contact

WeChat Official Account