Heteroduplex strand-specificity in restriction-stimulated recombination by the RecE pathway of Escherichia coli.
The RecE recombination pathway is active in Escherichia coli recB recC sbcA mutants. To isolate and characterize products and intermediates of RecE-mediated, break-induced, intramolecular recombination, we infected recB recC sbcA mutants, expressing EcoRI endonuclease, with chimeric lambda phages that allow EcoRI-mediated release of cloned linear recombination substrates. Substrates with direct terminal repeats recombined to yield a circular product with one copy of the repeated sequence. Some recombinants were heteroallelic for the recombining markers. Markers distant to the break were recovered in the circular product at a higher frequency than markers close to the break. To examine the heteroduplex structures that may have yielded the heteroallelic recombinants, nonreplicative substrates were employed. Some of the nonreplicative recombination products contained heteroduplexes, with a strong bias for paired strands ending 3' at the break. This strand bias in heteroduplex formation is consistent with recombination models that postulate homologous pairing of protruding 3' single-stranded ends.
Department of Molecular Genetics, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Z
Z
Silberstein
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M
Shalit
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A
Cohen
D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005819:Genetic Markers; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D009692:Nucleic Acid Heteroduplexes / Q000235:genetics*; D010957:Plasmids; D011995:Recombination, Genetic
D004269:DNA, Bacterial; D005819:Genetic Markers; D009692:Nucleic Acid Heteroduplexes; D004262:DNA Restriction Enzymes