Transcriptional and Mutational Analyses of theStreptomyces lividans recXGene and Its Interference with RecA Activity

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Abstract
The role of the 20,922-Da RecX protein and its interference with RecA activity were analyzed in Streptomyces lividans. TherecX gene is located 220 bp downstream of recA. Transcriptional analysis by reverse transcriptase PCR demonstrated thatrecX and recA constitute an operon. WhilerecA was transcribed at a basal level even under noninducing conditions, a recA-recX cotranscript was only detectable after induction of recA following DNA damage. The recA-recX cotranscript was less abundant than therecA transcript alone. The recX gene was inactivated by gene replacement. The resulting mutant had a clearly diminished colony size, but was not impaired in recombination activity, genetic instability, and resistance against UV irradiation. Expression of an extra copy of the S. lividans recA gene under control of the thiostrepton-inducible tipA promoter was lethal to the recX mutant, demonstrating that RecX is required to overcome the toxic effects of recA overexpression. Since inactivation of the recX gene did not influence transcription of recA, the putative function of the RecX protein might be the downregulation of RecA activity by interaction with the RecA protein or filament.