A combination of RNase H (rnh) and recBCD or sbcB mutations in Escherichia coli K12 adversely affects growth.
Colony forming ability of Escherichia coli strains carrying the rnh-339::cat mutant allele is strongly dependent on the recBCD and sbcB genes. A mutation inactivating either the RecBCD nuclease or exonuclease I (sbcB) is sufficient to restrict severely the efficiency of plating of strains carrying the rnh-339::cat mutation. Combining a non-lethal temperature-sensitive mutation in the RecBCD nuclease, recB270 (Ts) or recC271 (Ts), with rnh-339::cat renders strains temperature sensitive for growth, even though rnh+ strains with the recB270 (Ts) or recC271 (Ts) alleles are viable at 42 degrees C. The recombinational functions of the RecBCD nuclease can be excluded as the source of lethality on the basis of the following observations. Introduction of a recombination proficient, exonuclease defective recD1009 allele or production of the phage lambda GamS protein (an inhibitor of the RecBCD exonuclease activity) in an rnh-339::cat strain dramatically delays or impairs the ability of such strains to form colonies. Restoration of recombination proficiency by inclusion of an sbcB15 mutation with recB21 recC22 mutations does not restore the ability of the rnh-339::cat mutant strains to plate normally. A recBCD+ strain bearing the rnh-339::cat and sbcB15 mutations forms very few visible colonies after 24 h but forms colonies at normal frequencies after 48 h of incubation. Finally, plating efficiencies of strains are unaffected when the RecBCD recombination pathway is inactivated by introduction of recA56 into an rnh-339::cat strain. These results imply that the defective growth of rnh-339::cat recBCD strains is due to a defect in repair and not recombination mediated by either the RecBCD or the RecF pathway.
Itaya
M
M
Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
Crouch
R J
RJ
D000483:Alleles; D015139:Blotting, Southern; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004722:Endoribonucleases / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000235:genetics*; D005798:Genes, Bacterial; D005804:Genes, Lethal; D009154:Mutation; D011995:Recombination, Genetic; D016914:Ribonuclease H; D013696:Temperature
D004269:DNA, Bacterial; D015500:Chloramphenicol O-Acetyltransferase; D004722:Endoribonucleases; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V; D016914:Ribonuclease H