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PMID: 6090709 Published · ppublish English Journal Article

Selective protection of 5' ... GGCC ... 3' and 5' ... GCNGC ... 3' sequences by the hypermodified oxopyrimidine in Bacillus subtilis bacteriophage SP10 DNA.

Journal of virology ·Vol. 52 ·No. 1 ·1984-10-00 ·Pages 47-54

Wiatr CL, Witmer HJ

Abstract

The DNA of Bacillus subtilis bacteriophage SP10 is partially resistant to cleavage and methylation in vitro by restriction enzyme R . BsuRI and its cognate methylase even though greater than 20 copies of the target sequence, 5' ... GGCC ... 3', are present on the phage genome. YThy, a hypermodified oxopyrimidine that replaces a fraction of the thymine residues in SP10 DNA, was responsible for this protection, since YThy-free DNA was no longer resistant. Sites that were normally resistant could nevertheless be cleaved or methylated in vitro if the salt concentration was reduced or dimethyl sulfoxide was added to the reaction buffer. Analysis of the termini produced by cleavage suggested that resistant sites occurred in the sequence 5' ... GGCC-YThy ... 3', whereas sensitive sites, of which there were only two per genome, occurred in the sequence 5' ... GGCCG ... 3'. These in vitro results provide an explanation for the in vivo resistance of SP10 to restriction-modification by B. subtilis R. They also suggest ways in which the presence of the atypical base YThy in regions that flank the target might upset critical DNA-enzyme interactions necessary to locate and recognize the specific site of cleavage or methylation. YThy also strongly protected 5' ... GCNGC ... 3' (R . Fnu4HI) sequences on SP10 DNA, but the biological relevance of this protection is unclear.

MeSH Terms
Bacillus subtilis/analysis Bacteriophages/analysis Base Sequence DNA (Cytosine-5-)-Methyltransferases/metabolism DNA Restriction Enzymes/metabolism DNA, Viral/isolation & purification Genes, Viral Kinetics Methylation Oligodeoxyribonucleotides/analysis Substrate Specificity Thymine/analogs & derivatives,analysis
Chemicals
DNA, Viral Oligodeoxyribonucleotides DNA (Cytosine-5-)-Methyltransferases DNA Restriction Enzymes Thymine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Wiatr C L
Witmer H J
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39 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.52.1.47-54.1984
Published
1984-10-00
Pages
47-54
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC254487
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.52.1.47-54.1984 PMC Full Text View journal details
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