TY - JOUR
T1 - Structural and functional characterization of the recR gene of Streptomyces
AU - Peláez, A.
AU - Ribas-Aparicio, R.
AU - Gómez, A.
AU - Rodicio, M.
N1 - Funding Information:
Acknowledgements We thank Dr. H. Kieser and Prof. D. A. Hopwood (John Innes Institute, Norwich, England) for the gift of cosmids 3H4 and H66. R.M.R-A was the recipient of a fellowship from COFAA-BEDD from IPN and CONACyT, México. A.G. was recipient of a grant from FYCIT, Asturias, Spain. The work was supported by grant PB97-1074. DGICYT. Spain.
PY - 2001
Y1 - 2001
N2 - The recR gene product is necessary for homologous recombination and recombinational DNA repair in eubacteria. We report the isolation and sequencing of the recR gene from Streptomyces coelicolor. It encodes a protein of 198 amino acids, with a predicted molecular mass of 22 kDa. The deduced amino acid sequence shows significant similarity to that of RecR proteins from other bacteria, including Escherichia coli and Bacillus subtilis. Like these, Streptomyces RecR contains potential helix-hairpin-helix, zinc finger and A TP-binding motifs, as well as the Toprim domain which is present also in topoisomerases of Types IA and II, primases and nucleases of the OLD family. The recR genes of Escherichia coli and Bacillus subtilis are immediately preceded by a small ORF (orf12 and orf107, respectively). An equivalent ORF (orf1) is also found in Streptomyces. S. lividans recR mutants, obtained either by insertional inactivation of recR or by deletion of the gene together with the preceding ORF, displayed increased sensitivity to DNA-damaging agents (such as UV light and methylmethanesulfonate), when compared with the wild-type strain. Both mutants could be complemented by the wild-type orf1recR genes and also by the recR gene alone. Based on these results, orf1 appears to be dispensable for the repair function of Streptomyces RecR. In studies of heterologous complementation, the B. subtilis recR region (orf107recR) was found to complement the S. lividans Δorf1recR mutant, but the equivalent region from E. coli (orf12recR) could not. However, in the absence of orf107, B. subtilis recR was unable to restore the wild-type phenotype to the Streptomyces deletion mutant.
AB - The recR gene product is necessary for homologous recombination and recombinational DNA repair in eubacteria. We report the isolation and sequencing of the recR gene from Streptomyces coelicolor. It encodes a protein of 198 amino acids, with a predicted molecular mass of 22 kDa. The deduced amino acid sequence shows significant similarity to that of RecR proteins from other bacteria, including Escherichia coli and Bacillus subtilis. Like these, Streptomyces RecR contains potential helix-hairpin-helix, zinc finger and A TP-binding motifs, as well as the Toprim domain which is present also in topoisomerases of Types IA and II, primases and nucleases of the OLD family. The recR genes of Escherichia coli and Bacillus subtilis are immediately preceded by a small ORF (orf12 and orf107, respectively). An equivalent ORF (orf1) is also found in Streptomyces. S. lividans recR mutants, obtained either by insertional inactivation of recR or by deletion of the gene together with the preceding ORF, displayed increased sensitivity to DNA-damaging agents (such as UV light and methylmethanesulfonate), when compared with the wild-type strain. Both mutants could be complemented by the wild-type orf1recR genes and also by the recR gene alone. Based on these results, orf1 appears to be dispensable for the repair function of Streptomyces RecR. In studies of heterologous complementation, the B. subtilis recR region (orf107recR) was found to complement the S. lividans Δorf1recR mutant, but the equivalent region from E. coli (orf12recR) could not. However, in the absence of orf107, B. subtilis recR was unable to restore the wild-type phenotype to the Streptomyces deletion mutant.
KW - Genetic complementation
KW - RecR
KW - Recombinational DNA repair
KW - Streptomyces coelicolor
KW - Streptomyces lividans
UR - http://www.scopus.com/inward/record.url?scp=0034888480&partnerID=8YFLogxK
U2 - 10.1007/s004380100460
DO - 10.1007/s004380100460
M3 - Artículo
SN - 1617-4615
VL - 265
SP - 663
EP - 672
JO - Molecular Genetics and Genomics
JF - Molecular Genetics and Genomics
IS - 4
ER -