Conservation of RecG activity from pathogens to hyperthermophiles.
Maintaining the integrity of the genome is essential for the survival of all organisms. RecG helicase plays an important part in this process in Escherichia coli, promoting recombination and DNA repair, and providing ways to rescue stalled replication forks by way of a Holliday junction intermediate. We purified RecG proteins from three other species: two Gram-positive mesophiles, Bacillus subtilis and Streptococcus pneumoniae, and one extreme thermophile, Aquifex aeolicus. All three proteins bind and unwind replication fork and Holliday junction DNA molecules with efficiencies similar to the E. coli protein. Proteins from the Gram-positive species promote DNA repair in E. coli, indicating either that RecG acts alone or that any necessary protein-protein interactions are conserved. The S. pneumoniae RecG reduces plasmid copy number when expressed in E. coli, indicating that like the E. coli protein it unwinds plasmid R loop structures used to prime replication. This effect is not seen with B. subtilis RecG; the protein either lacks R loop unwinding activity or is compromised by having insufficient ATP. The A. aeolicus protein unwinds DNA well at 60 degrees C but is less efficient at 37 degrees C, explaining its inability to function in E. coli at this temperature. The N-terminal extension present in this protein was investigated and found to be dispensable for activity and thermo-stability. The results presented suggest that the role of RecG in DNA replication and repair is likely to be conserved throughout all bacteria, which underlines the importance of this protein in genome duplication and cell survival.
Wen
Qin
Q
Queens Medical Centre, Institute of Genetics, University of Nottingham, Nottingham NG7 2UH, UK.
Mahdi
Akeel A
AA
Briggs
Geoffrey S
GS
Sharples
Gary J
GJ
Lloyd
Robert G
RG
D001419:Bacteria / Q000235:genetics* / Q000528:radiation effects; D020029:Base Pairing; D001483:Base Sequence; D019295:Computational Biology; D004265:DNA Helicases / Q000235:genetics / Q000502:physiology*; D004260:DNA Repair; D004261:DNA Replication / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay; D029968:Escherichia coli Proteins / Q000235:genetics / Q000502:physiology*; D015870:Gene Expression; D017385:Sequence Homology; D013045:Species Specificity; D013696:Temperature; D014466:Ultraviolet Rays
D029968:Escherichia coli Proteins; C071446:RecG protein, E coli; D004265:DNA Helicases