Cleavage of methylated CCCGGG sequences containing either N4-methylcytosine or 5-methylcytosine with MspI, HpaII, SmaI, XmaI and Cfr9I restriction endonucleases.

Nucleic acids research ·Vol. 15 ·No. 17 ·1987-09-11 ·Pages 7091-102

Butkus V, Petrauskiene L, Maneliene Z, Klimasauskas S, Laucys V, Janulaitis A

Abstract

The cleavage specificity of R.Cfr9I was determined to be C decreases CCGGG whereas the methylation specificity of M.Cfr9I was C4mCCGGG. The action of MspI, HpaII, SmaI, XmaI and Cfr9I restriction endonucleases on an unmethylated parent d(GGACCCGGGTCC) dodecanucleotide duplex and a set of oligonucleotide duplexes, containing all possible substitutions of either 4mC or 5mC for C in the CCCGGG sequence, was investigated. It was found that 4mC methylation, in contrast to 5mC, renders the CCCGGG site resistant to practically all the investigated endonucleases. The cleavage of methylated substrates with restriction endonucleases is discussed.

MeSH Terms

5-Methylcytosine Base Sequence Cytosine/analogs & derivatives,analysis DNA Restriction Enzymes/metabolism Nucleotide Mapping Oligodeoxyribonucleotides Substrate Specificity

Chemicals

Oligodeoxyribonucleotides N(4)-methylcytosine 5-Methylcytosine Cytosine DNA Restriction Enzymes

Authors & Affiliations

6 authors, click to expand affiliations / ORCID

Butkus V

ESP Fermentas, All Union Research Institute of Applied Enzymology, Lithuanian SSR.

Petrauskiene L

Maneliene Z

Klimasauskas S

Laucys V

Janulaitis A

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17 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/15.17.7091

Published

1987-09-11

Pages

7091-102

Language

English

Region

England

NLM ID

0411011

PMCID

PMC306195

Subset

External Links

PubMed source DOI: 10.1093/nar/15.17.7091 PMC Full Text View journal details

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Cleavage of methylated CCCGGG sequences containing either N4-methylcytosine or 5-methylcytosine with MspI, HpaII, SmaI, XmaI and Cfr9I restriction endonucleases.

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