The Bacillus subtilis AddAB helicase/nuclease is regulated by its cognate Chi sequence in vitro

Frederic Chedin, S. Dusko Ehrlich, Stephen C. Kowalczykowski

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

The AddAB enzyme is important to homologous DNA recombination in Bacillus subtilis, where it is thought to be the functional counterpart of the RecBCD enzyme of Escherichi a coli. In vivo, AddAB responds to a specific five-nucleotide sequence (5'-AGCGG-3' or its complement) in a manner analogous to the response of the RecBCD enzyme to interaction with χ sequences. Here, we show that purified AddAB enzyme is able to load at a double-stranded DNA end and is both a DNA helicase and nuclease, whose combined action results in the degradation of both strands of the DNA duplex. During translocation, recognition of the properly oriented sequence 5'-AGCGG-3' causes attenuation of the AddAB enzyme nuclease activity that is responsible for degradation of the strand 3'-terminal at the entry site. Therefore, we conclude that 5'-AGCGG-3' is the B. subtilis Chi site and it is hereafter referred to as χ(Bs). After encountering χ(Bs), both the degradation of the 5'-terminal strand and the helicase activity persist. Thus, processing of a double-stranded DNA end by the AddAB enzyme produces a duplex DNA molecule with a protruding 3'-terminated single-stranded tail, a universal intermediate of the recombination process. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)7-20
Number of pages14
JournalJournal of Molecular Biology
Volume298
Issue number1
DOIs
StatePublished - Apr 21 2000

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Bacillus subtilis
Exodeoxyribonuclease V
DNA
DNA Helicases
Deoxyribonucleases
Homologous Recombination
Genetic Recombination
In Vitro Techniques
AddAB enzyme

Keywords

  • AddAB
  • Chi
  • Helicase
  • Homologous recombination
  • Nuclease

ASJC Scopus subject areas

  • Virology

Cite this

The Bacillus subtilis AddAB helicase/nuclease is regulated by its cognate Chi sequence in vitro. / Chedin, Frederic; Ehrlich, S. Dusko; Kowalczykowski, Stephen C.

In: Journal of Molecular Biology, Vol. 298, No. 1, 21.04.2000, p. 7-20.

Research output: Contribution to journalArticle

Chedin, Frederic ; Ehrlich, S. Dusko ; Kowalczykowski, Stephen C. / The Bacillus subtilis AddAB helicase/nuclease is regulated by its cognate Chi sequence in vitro. In: Journal of Molecular Biology. 2000 ; Vol. 298, No. 1. pp. 7-20.
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