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doi:10.1534/genetics.105.052076
A more recent version of this article appeared on April 1, 2006.
REGULAR RESEARCH PAPERS |
Functions of multiple exonucleases are essential for cell viability, DNA repair and homologous recombination in recD mutants of Escherichia coli
Damir Dermic 1*
1 Ruder Boskovic Institute
* To whom correspondence should be addressed. E-mail: dermic{at}irb.hr.
Submitted on October 6, 2005
Revised on November 18, 2005
Accepted on 18 January 2006
Heterotrimeric RecBCD enzyme unwinds and resects DNA duplex containing blunt double strand ends, and directs loading of strand-exchange protein RecA onto the unwound 3'-ending strand, thereby initiating the majority of recombination in wild-type Escherichia coli. When the enzyme lacks its RecD subunit, the resulting RecBC enzyme, active in recD mutants, is recombination proficient although it has just helicase and RecA loading activity, but is not a nuclease. However, E. coli encodes for several other exonucleases that digest double-stranded and single-stranded DNA, and thus might act in consort with the RecBC enzyme to efficiently promote recombination reactions. To test this hypothesis, I inactivated multiple exonucleases (i.e. exonuclease I, exonuclease X, exonuclease VII, RecJ, SbcCD) in recD derivatives of wild-type and nuclease-deficient recB1067 strain and assessed the ability of the resultant mutants to maintain cell viability, and promote DNA repair and homologous recombination. A complex pattern of overlapping and sometimes competing activities of multiple exonucleases in recD mutants was thus revealed. These exonucleases were shown to be essential for cell viability, DNA repair (of UV and ã induced lesions) and homologous recombination (during Hfr conjugation and P1 transduction), dependent on RecBC enzyme. A model for donor DNA processing in recD transconjugants and transductants was proposed.
Key Words: DNA double-strand break repair, Hfr conjugation, P1 transduction, RecBC enzyme, exonucleases: ExoI, ExoVII, ExoX, RecJ, SbcCD
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