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PMID: 8393986 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

An E. coli RuvC mutant defective in cleavage of synthetic Holliday junctions.

Nucleic acids research ·Vol. 21 ·No. 15 ·1993-07-25 ·Pages 3359-64

Sharples GJ, Lloyd RG

Abstract

Escherichia coli RuvC protein is a specific endonuclease that resolves recombination intermediates into viable products. The structural features needed for RuvC activity were investigated by sequencing three ruvC mutations and relating the base pair changes identified to the activity of the mutant proteins. Each of the three mutations is a single base-pair substitution. ruvC51 converts glycine-15 to an aspartic acid residue. The product of ruvC51 was purified and shown to retain the ability to bind junctions, albeit with a slightly reduced affinity. However, it has lost the ability to resolve these structures by symmetrical cleavage. A multicopy ruvC51 plasmid confers sensitivity to UV light in a ruvC+ strain. The ruvC53 allele causes a glycine-17 to serine substitution while ruvC55 produces a stop codon. Neither of these genes produces a stable product. The results suggest that the N-terminal domain of RuvC may be concerned with cleavage of junctions.

Related Genes
ruvC
MeSH Terms
Amino Acid Sequence Bacterial Proteins/chemistry,genetics,metabolism Base Sequence Cloning, Molecular Codon Endodeoxyribonucleases Escherichia coli/genetics,radiation effects Escherichia coli Proteins Glycine Molecular Sequence Data Mutation Nucleotidyltransferases/metabolism Plasmids Polymerase Chain Reaction Rec A Recombinases/metabolism Recombinant Proteins/metabolism Serine Structure-Activity Relationship Transposases Ultraviolet Rays
Chemicals
Bacterial Proteins Codon Escherichia coli Proteins Recombinant Proteins ruvC protein, E coli Serine Nucleotidyltransferases Rec A Recombinases Transposases Endodeoxyribonucleases Glycine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Sharples G J
Department of Genetics, University of Nottingham, Queens Medical Centre, UK.
Lloyd R G
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41 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/21.15.3359
Published
1993-07-25
Pages
3359-64
Language
English
Region
England
NLM ID
0411011
PMCID
PMC331431
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1093/nar/21.15.3359 PMC Full Text View journal details
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