PriA mutations that affect PriA-PriC function during replication restart.
In Escherichia coli, repair and restart of collapsed replication forks is thought to be essential for cell growth. The replication restart proteins, PriA, PriB, PriC, DnaB, DnaC, DnaG, DnaT and Rep, form redundant pathways that recognize repaired replication forks and restart them. Recognition, modulation of specific DNA structures and loading of the replicative helicase by the replication restart proteins, is likely to be important for replication restart. It has been hypothesized that PriB and PriC function with PriA in genetically separate and redundant PriA-PriB and PriA-PriC pathways. In this study, the del(priB)302 or priC303:kan mutations were used to isolate the PriA-PriB and PriA-PriC pathways genetically so that the effects of three priA missense mutations, priA300 (K230R), priA301 (C479Y) and priA306 (L557P), on these pathways could be assessed. In a wild-type background, the three priA mutations had little, if any, effect on the phenotypes of UV resistance, basal levels of SOS expression and cell viability. In the priB mutant, priA300 and priA301 caused dramatic negative changes in the three phenotypes listed above (and others), whereas the third priA mutant allele, priA306, showed very little negative effect. In the priC mutant, all three priA mutations behaved similarly, producing little, if any, changes in phenotypes. We conclude that priA300 and priA301 mostly affect the PriA-PriC pathway and do so more than priA306. We suggest that PriA's helicase activity is important for the PriA-PriC pathway of replication restart.
Sandler
S J
SJ
203 Morrill Science Center IVN, Department of Microbiology, University of Massachusetts, Amherst, MA 01003, USA. sandler@microbio.umass.edu
McCool
J D
JD
Do
T T
TT
Johansen
R U
RU
D001426:Bacterial Proteins / Q000378:metabolism*; D010583:Bacteriophage mu / Q000502:physiology; D002455:Cell Division / Q000528:radiation effects; D004261:DNA Replication / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000528:radiation effects / Q000821:virology; D029968:Escherichia coli Proteins; D005804:Genes, Lethal / Q000235:genetics; D005838:Genotype; D008954:Models, Biological; D020125:Mutation, Missense / Q000235:genetics*; D010641:Phenotype; D051716:Replication Protein A; D013014:SOS Response, Genetics / Q000235:genetics; D014466:Ultraviolet Rays
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D051716:Replication Protein A; C069996:PriB protein (N-terminal); C069995:PriC protein, E coli