Replicase, excisionase, and integrase genes of the Streptomyces element pSAM2 constitute an operon positively regulated by the pra gene.
pSAM2 is a site-specific integrative element from Streptomyces ambofaciens. The pra gene described earlier as an activator of pSAM2 replication is shown here to be also involved in the activation of its integration and excision. This was evidenced with derivatives of pSAM2 mutant B3 in which the pra gene was placed under the control of the inducible tipAp promoter. Transformation of Streptomyces lividans by these derivatives was efficient only when pra expression was induced, indicating its involvement in pSAM2 integration activation. Once established, these constructions remained integrated in the chromosome under noninduced conditions. Activation of the pra expression provoked strong activation of their excision, leading to the appearance of free forms. The results of functional, transcriptional, and sequence analyses allowed to conclude that the three genes repSA, xis, and int coding for the pSAM2 replicase, excisionase, and integrase, respectively, constitute an operon directly or indirectly activated by pra.
Sezonov
G
G
Laboratoire de Biologie et Génétique Moléculaire, URA CNRS 2225, Université Paris-Sud, Orsay, France. sezonov@igmors.u-psud.fr
Duchêne
A M
AM
Friedmann
A
A
Guérineau
M
M
Pernodet
J L
JL
D001483:Base Sequence; D002874:Chromosome Mapping; D004265:DNA Helicases / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D005798:Genes, Bacterial; D019426:Integrases / Q000235:genetics*; D008969:Molecular Sequence Data; D009876:Operon; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics; D014158:Transcription, Genetic; D014170:Transformation, Genetic; D014764:Viral Proteins
D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D015534:Trans-Activators; D014764:Viral Proteins; C045264:replication initiator protein; D004254:DNA Nucleotidyltransferases; C023140:DNA replicase; D019426:Integrases; C063911:excisionase; D004259:DNA-Directed DNA Polymerase; D004265:DNA Helicases