The reduced levels of χ recognition exhibited by the RecBC1004D enzyme reflect its recombination defect in vivo

D. A. Arnold, P. R. Bianco, S. C. Kowalczykowski

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

Homologous recombination in Escherichia coli is initiated by the RecBCD enzyme and is stimulated by an 8-nucleotide element known as Chi (χ). We present a detailed biochemical characterization of a mutant RecBCD enzyme, designated RecBC1004D, that displays a reduced level of χ site recognition. Initially characterized genetically as unable to respond to the χ sequence, we provide evidence to indicate that the ability of this mutant enzyme to respond to χ is reduced rather than lost; the mutant displays about 20-fold lower χ recognition than wild-type RecBCD enzyme. Although this enzyme exhibits wild-type levels of double-stranded DNA exonuclease, helicase, and ATPase activity, its ability to degrade single-stranded DNA is enhanced 2-3-fold. The data presented here suggest that the reduced recombination proficiency of the recBC1004D strain observed in vivo results from a basal level of modification of the RecBC1004D enzyme at both χ-specific, as well as nonspecific, DNA sequences.

Original languageEnglish (US)
Pages (from-to)16476-16486
Number of pages11
JournalJournal of Biological Chemistry
Volume273
Issue number26
DOIs
StatePublished - Jun 26 1998

ASJC Scopus subject areas

  • Biochemistry

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