NDLI: Deletion of One Nucleotide within the Homonucleotide Tract Present in the hsdS Gene Alters the DNA Sequence Specificity of Type I Restriction-Modification System NgoAV ▿ †

Content Provider	PubMed Central
Author	Monika, Adamczyk-poplawska Lower, Michal Piekarowicz, Andrzej
Copyright Year	2011
Abstract	As a result of a frameshift mutation, the hsdS locus of the NgoAV type IC restriction and modification (RM) system comprises two genes, hsdS NgoAV1 and hsdS NgoAV2. The specificity subunit, HsdSNgoAV, the product of the hsdS NgoAV1 gene, is a naturally truncated form of an archetypal specificity subunit (208 N-terminal amino acids instead of 410). The presence of a homonucleotide tract of seven guanines (poly[G]) at the 3′ end of the hsdS NgoAV1 gene makes the NgoAV system a strong candidate for phase variation, i.e., stochastic addition or reduction in the guanine number. We have constructed mutants with 6 guanines instead of 7 and demonstrated that the deletion of a single nucleotide within the 3′ end of the hsdS NgoAV1 gene restored the fusion between the hsdS NgoAV1 and hsdS NgoAV2 genes. We have demonstrated that such a contraction of the homonucleotide tract may occur in vivo: in a Neisseria gonorrhoeae population, a minor subpopulation of cells appeared to have only 6 guanines at the 3′ end of the hsdS NgoAV1 gene. Escherichia coli cells carrying the fused gene and expressing the NgoAVΔ RM system were able to restrict λ phage at a level comparable to that for the wild-type NgoAV system. NgoAV recognizes the quasipalindromic interrupted sequence 5′-GCA(N8)TGC-3′ and methylates both strands. NgoAVΔ recognizes DNA sequences 5′-GCA(N7)GTCA-3′ and 5′-GCA(N7)CTCA-3′, although the latter sequence is methylated only on the complementary strand within the 5′-CTCA-3′ region of the second recognition target sequence.
Related Links	http://dx.doi.org/10.1128/jb.05672-11
Ending Page	6759
Page Count	10
Starting Page	6750
File Format	PDF
ISSN	00219193
e-ISSN	10985530
Journal	Journal of Bacteriology
Issue Number	23
Volume Number	193
Language	English
Publisher	American Society for Microbiology
Publisher Date	2011-12-01
Access Restriction	Open
Rights Holder	American Society for Microbiology
Subject Keyword	Molecular Biology Microbiology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Microbiology

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Deletion of One Nucleotide within the Homonucleotide Tract Present in the hsdS Gene Alters the DNA Sequence Specificity of Type I Restriction-Modification System NgoAV ▿ †

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