Construction of recB-recD genetic fusion and functional analysis of RecBDC fusion enzyme in Escherichia coli.

ABSTRACT:

Background

recD, located between recB and argA, encodes the smallest polypeptide (60 kDa) of the heterotrimeric enzyme RecBCD in Escherichia coli. RecD is a 5'-3' helicase and is required for the nuclease activity of RecBCD and for tight binding to dsDNA ends. Here, we have tested the hypothesis that RecD regulates the structure and activities of RecBCD, including RecA loading.

Results

To characterize its regulatory functions, recD was genetically fused to recB through deletion and substitution mutations. The recB-recD fusion led to a decreased amount of the heterotrimer. Both fusion mutants proved to be recombination proficient, viable and resistant to DNA damaging agents, and to have DNA unwinding, ATP-dependent dsDNA exonuclease and Chi genetic activities.

Conclusion</

SUBMITTER: Portakal O 

PROVIDER: S-EPMC2586629 | biostudies-literature | 2008 Oct

REPOSITORIES: biostudies-literature