Activation of Chi recombinational hotspots by RecBCD-like enzymes from enteric bacteria.
Chi sites, 5'G-C-T-G-G-T-G-G-3', enhance homologous recombination in Escherichia coli and are activated by the RecBCD enzyme. To test the ability of Chi to be activated by analogous enzymes from other bacteria, we cloned recBCD-like genes from diverse bacteria into an E. coli recBCD deletion mutant. Clones from seven species of enteric bacteria conferred to this deletion mutant recombination proficiency, Chi hotspot activity in lambda Red- Gam- vegetative crosses, and RecBCD enzyme activities, including Chi-dependent DNA strand cleavage. Three clones from Pseudomonas aeruginosa and Ps. putida conferred recombination proficiency and ATP-dependent nuclease activity, but neither Chi hotspot activity nor Chi-dependent DNA cleavage. These results imply that Chi has been conserved as a recombination-promoting signal for RecBCD-like enzymes in enteric bacteria but not in more distantly related bacteria such as Pseudomonas spp. We discuss the possibility that other, presently unknown, nucleotide sequences serve the same function as Chi in Pseudomonas spp.
McKittrick
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Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
Smith
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D000255:Adenosine Triphosphate / Q000378:metabolism; D010582:Bacteriophage lambda / Q000235:genetics; D003001:Cloning, Molecular; D004265:DNA Helicases / Q000235:genetics; D004755:Enterobacteriaceae / Q000235:genetics*; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D005816:Genetic Complementation Test; D010957:Plasmids; D011995:Recombination, Genetic; D010948:Viral Plaque Assay; D014779:Virus Replication
D000255:Adenosine Triphosphate; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V; D004265:DNA Helicases