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PMID:25246477
| Citation |
Qin, W, Liu, NN, Wang, L, Zhou, M, Ren, H, Bugnard, E, Liu, JL, Zhang, LH, Vendôme, J, Hu, JS and Xi, XG (2014) Characterization of biochemical properties of Bacillus subtilis RecQ helicase. J. Bacteriol. 196:4216-28 |
|---|---|
| Abstract |
RecQ family helicases function as safeguards of the genome. Unlike Escherichia coli, the Gram-positive Bacillus subtilis bacterium possesses two RecQ-like homologues, RecQ[Bs] and RecS, which are required for the repair of DNA double-strand breaks. RecQ[Bs] also binds to the forked DNA to ensure a smooth progression of the cell cycle. Here we present the first biochemical analysis of recombinant RecQ[Bs]. RecQ[Bs] binds weakly to single-stranded DNA (ssDNA) and blunt-ended double-stranded DNA (dsDNA) but strongly to forked dsDNA. The protein exhibits a DNA-stimulated ATPase activity and ATP- and Mg(2+)-dependent DNA helicase activity with a 3' → 5' polarity. Molecular modeling shows that RecQ[Bs] shares high sequence and structure similarity with E. coli RecQ. Surprisingly, RecQ[Bs] resembles the truncated Saccharomyces cerevisiae Sgs1 and human RecQ helicases more than RecQ[Ec] with regard to its enzymatic activities. Specifically, RecQ[Bs] unwinds forked dsDNA and DNA duplexes with a 3'-overhang but is inactive on blunt-ended dsDNA and 5'-overhung duplexes. Interestingly, RecQ[Bs] unwinds blunt-ended DNA with structural features, including nicks, gaps, 5'-flaps, Kappa joints, synthetic replication forks, and Holliday junctions. We discuss these findings in the context of RecQ[Bs]'s possible functions in preserving genomic stability. |
| Links |
PubMed PMC4248853 Online version:10.1128/JB.06367-11 |
| Keywords |
Adenosine Triphosphatases/metabolism; Adenosine Triphosphate/metabolism; Bacillus subtilis/enzymology; DNA/metabolism; DNA, Single-Stranded/metabolism; Magnesium/metabolism; Models, Molecular; Protein Binding; RecQ Helicases/metabolism; Recombinant Proteins/genetics; Recombinant Proteins/isolation & purification; Recombinant Proteins/metabolism |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0043138: 3'-5' DNA helicase activity |
ECO:0000314: |
F |
Figure 5. show RecQ is a 3'to5' DNA helicase. It unwind NDA substrate with 3' single stranded region. |
complete | ||||
Notes
See also
References
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