RecA433 cells are defective in recF-mediated processing of disrupted replication forks but retain recBCD-mediated functions.
RecA is required for recombinational processes and cell survival following UV-induced DNA damage. recA433 is a historically important mutant allele that contains a single amino acid substitution (R243H). This mutation separates the recombination and survival functions of RecA. recA433 mutants remain proficient in recombination as measured by conjugation or transduction, but are hypersensitive to UV-induced DNA damage. The cellular functions carried out by RecA require either recF pathway proteins or recBC pathway proteins to initiate RecA-loading onto the appropriate DNA substrates. In this study, we characterized the ability of recA433 to carry out functions associated with either the recF pathway or recBC pathway. We show that several phenotypic deficiencies exhibited by recA433 mutants are similar to recF mutants but distinct from recBC mutants. In contrast to recBC mutants, recA433 and recF mutants fail to process or resume replication following disruption by UV-induced DNA damage. However, recA433 and recF mutants remain proficient in conjugational recombination and are resistant to formaldehyde-induced protein-DNA crosslinks, functions that are impaired in recBC mutants. The results are consistent with a model in which the recA433 mutation selectively impairs RecA functions associated with the RecF pathway, while retaining the ability to carry out RecBCD pathway-mediated functions. These results are discussed in the context of the recF and recBC pathways and the potential substrates utilized in each case.
Al-Hadid
Qais
Q
Department of Biology, Portland State University, Portland, OR 97207, USA. qalhadid@uci.edu
Ona
Katherine
K
Courcelle
Charmain T
CT
Courcelle
Justin
J
D019943:Amino Acid Substitution; D003432:Cross-Linking Reagents; D004249:DNA Damage; D004261:DNA Replication; D004269:DNA, Bacterial / Q000187:drug effects / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism*; D043211:Exodeoxyribonuclease V / Q000235:genetics / Q000378:metabolism*; D005557:Formaldehyde; D005798:Genes, Bacterial; D008954:Models, Biological; D009154:Mutation; D011938:Rec A Recombinases / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; C489340:recF protein, E coli; D005557:Formaldehyde; D011938:Rec A Recombinases; D043211:Exodeoxyribonuclease V; C468322:exodeoxyribonuclease V, E coli