Recombination and annealing pathways compete for substrates in making rrn duplications in Salmonella enterica.
Tandem genetic duplications arise frequently between the seven directly repeated 5.5-kb rrn loci that encode ribosomal RNAs in Salmonella enterica. The closest rrn genes, rrnB and rrnE, flank a 40-kb region that includes the purHD operon. Duplications of purHD arise by exchanges between rrn loci and form at a high rate (10(-3)/cell/division) that remains high in strains blocked for early steps in recombination (recA, recB, and/or recF), but drops 30-fold in mutants blocked for later Holliday junction resolution (ruvC recG). The duplication defect of a ruvC recG mutant was fully corrected by an added mutation in any one of the recA, recB, or recF genes. To explain these results, we propose that early recombination defects activate an alternative single-strand annealing pathway for duplication formation. In wild-type cells, rrn duplications form primarily by the action of RecFORA on single-strand gaps. Double-strand breaks cannot initiate rrn duplications because rrn loci lack Chi sites, which are essential for recombination between two separated rrn sequences. A recA or recF mutation allows unrepaired gaps to accumulate such that different rrn loci can provide single-strand rrn sequences that lack the RecA coating that normally inhibits annealing. A recB mutation activates annealing by allowing double-strand ends within rrn to avoid digestion by RecBCD and provide a new source of rrn ends for use in annealing. The equivalent high rates of rrn duplication by recombination and annealing pathways may reflect a limiting economy of gaps and breaks arising in heavily transcribed, palindrome-rich rrn sequences.
Department of Microbiology and Molecular Genetics, University of California, Davis, California 95616.
Andrew B
AB
Reams
Eric
E
Kofoid
Natalie
N
Duleba
John R
JR
Roth
D001426:Bacterial Proteins / Q000235:genetics; D002876:Chromosomes, Bacterial / Q000235:genetics*; D053903:DNA Breaks, Double-Stranded; D004265:DNA Helicases / Q000235:genetics; D053843:DNA Mismatch Repair / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D043211:Exodeoxyribonuclease V / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics*; D011938:Rec A Recombinases / Q000235:genetics; D019779:Salmonella enterica / Q000235:genetics*; D020080:Tandem Repeat Sequences / Q000235:genetics*; D012407:rRNA Operon / Q000235:genetics
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D012335:RNA, Ribosomal; C066218:recF protein, Bacteria; D011938:Rec A Recombinases; D043211:Exodeoxyribonuclease V; D004265:DNA Helicases
duplication; recombination; ribosomal RNA loci; single-strand annealing