Recombination and Annealing Pathways Compete for Substrates in Making rrn Duplications in Salmonella enterica. Issue 1 (1st January 2014)

• Record Type: Journal Article
• Authors: Reams, Andrew B; Kofoid, Eric; Duleba, Natalie; Roth, John R
• Is Part Of: Genetics. Volume 196:Issue 1(2014)
• Page Start: 119
• Page End: 135
• Publication Date: 2014-01-01
• Subjects: duplication -- recombination -- single-strand annealing -- ribosomal RNA loci; Genetics -- Periodicals; 576.5
• DOI: 10.1534/genetics.113.158519
• Languages: English
• ISSNs: 0016-6731

Abstract

Abstract: 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.