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Detection and Quantitation of RecBCD Enzyme (Exonuclease V) Activity

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DNA Repair Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 152))

abstract

The RecBCD enzyme serves two functions in the bacterial cell: it is a nuclease that destroys linear double-stranded DNA (dsDNA), and a DNA helicase that generates single-stranded DNA (ssDNA) used by RecA protein to initiate homologous recombination (1–3). A specific DNA sequence called Chi (5′-GCTGGTGG) is a signal that regulates these two functions (1,3). An encounter with Chi by RecBCD during its reaction with dsDNA leads to suppression of the nuclease activity and reveals the recombination-initiating function of RecBCD (1,3). The enzyme in Escherichia coli and other bacteria consists of three protein subunits encoded by the recB, recC, and recD genes, whereas some bacteria (e.g., Bacillus subtilis, Lactococcus lactis) produce a two-subunit enzyme [AddAB and RexAB, respectively (4,5)]. The genes encoding these latter enzymes complement an E. coli recBCD deletion mutation in vivo (5,6), and the purified AddAB enzyme has similar catalytic activity to the more extensively studied E. coli enzyme (7).

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Maryland, College Park, MD

    Douglas A. Julin

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  1. Douglas A. Julin
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  1. HiberGen Ltd. and University College, Cork, Ireland

    Pat Vaughan

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© 2000 Humana Press Inc.

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Julin, D.A. (2000). Detection and Quantitation of RecBCD Enzyme (Exonuclease V) Activity. In: Vaughan, P. (eds) DNA Repair Protocols. Methods in Molecular Biology™, vol 152. Humana Press. https://doi.org/10.1385/1-59259-068-3:91

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  • DOI: https://doi.org/10.1385/1-59259-068-3:91

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-643-7

  • Online ISBN: 978-1-59259-068-1

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