Influence of disruption of the recA gene on genetic instability and genome rearrangement in Streptomyces lividans.
Streptomyces lividans TK23 gives rise to chloramphenicol-sensitive (Cml(s)) mutants at a frequency of about 0.5%. This is due to the frequent occurrence of very large chromosomal deletions removing the corresponding chloramphenicol resistance gene. A mutant in which the recA gene has been disrupted (S. lividans FrecD3 [G. Muth, D. Frese, A. Kleber, and W. Wohlleben, personal communication]) segregated about 70 times more chloramphenicol-sensitive mutants than the parental strain. An enhancement of the deletion frequency was responsible for this mutator phenotype. The amplifiable locus AUD1 has a duplicated structure in some S. lividans strains and is frequently highly amplified in some mutants generated by genetic instability. The chromosomal AUD1 is not amplified in strain TK23 because of the lack of one duplication. Nevertheless, AUD1-derived amplifiable units presenting the typical duplicated organization amplified very well in TK23 when carried on a plasmid. No amplification of these units was observed in the recA mutant. The ability to amplify was restored when the wild-type recA gene was introduced into the plasmid carrying the amplifiable unit. These results suggest that the RecA protein plays a role in reducing the level of genetic instability and chromosomal deletions and show that the recA gene is necessary to achieve high-copy-number amplification of AUD1.
Volff
J N
JN
Institut für Industrielle Genetik, Universität Stuttgart, Germany.
Altenbuchner
J
J
D002876:Chromosomes, Bacterial / Q000648:ultrastructure; D004269:DNA, Bacterial / Q000648:ultrastructure; D005784:Gene Amplification; D015321:Gene Rearrangement; D005798:Genes, Bacterial; D011938:Rec A Recombinases / Q000235:genetics*; D011995:Recombination, Genetic; D017384:Sequence Deletion; D013302:Streptomyces / Q000235:genetics*
D004269:DNA, Bacterial; D011938:Rec A Recombinases