title: Strand targeting signal(s) for in vivo mutation avoidance by post-replication mismatch repair in Escherichia coli.
abstract: The involvement of GATC sites in directing mismatch correction for the elimination of replication errors in Escherichia coli was investigated in vivo by analyzing mutation rates for a gene carried on a series of related plasmids that contain 2, 1 and 0 such sites. This gene encoding chloramphenicol acetyl transferase (Cat protein) was inactivated by a point mutation. In vivo mutations restoring resistance to chloramphenicol were scored in mismatch repair proficient (mut+) and deficient (mutHLS-) strains. In mut+ cells, reduction of GATC sites from 2 to 0 increased mutation rates approximately 10-fold. Removal of the GATC site distal to the cat- mutation increased the rate of mutation less than 2-fold, indicating that mismatch repair can proceed normally with a single site. The mutation rate increased 3-fold after removal of the GATC site proximal to the mutation. In the absence of a GATC site, mutL- and mutS- strains exhibited a 2- to 3-fold increased mutation rate as compared to isogenic mutH- and mut+ strains. This indicates that 50%-70% of replication errors can be corrected in a mutLS-dependent way in the absence of any GATC site to target mismatch correction to newly synthesized DNA strands. Other strand targeting signals, possibly single strand discontinuities, might be used in mutLS-dependent repair.
authors.affiliation: Centre de Recherche de Biochimie et de Génétique Cellulaires du C.N.R.S., Université Paul Sabatier, Toulouse, France.
authors.forename: J P
authors.initials: JP
authors.lastname: Claverys
authors.forename: V
authors.initials: V
authors.lastname: Méjean
mesh_terms: D001482:Base Composition; D001483:Base Sequence; D002702:Chloramphenicol Resistance / Q000235:genetics; D004260:DNA Repair; D004261:DNA Replication; D004269:DNA, Bacterial / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics*; D009154:Mutation; D010957:Plasmids; D015183:Restriction Mapping; D014170:Transformation, Genetic
chemical_list: D004269:DNA, Bacterial