The rec locus, a competence-induced operon in Streptococcus pneumoniae.
To study competence and the process of transformation (TFN) in pneumococci, we developed a method for isolating TFN- mutants using insertional inactivation coupled with fusions to the gene for alkaline phosphatase (phoA). One TFN- mutant transformed 2 log units less efficiently than the parent strain. Reconstitution of the mutated region revealed a locus, rec, that contains two polycistronic genes, exp10 and the previously identified recA (B. Martin, J. M. Ruellan, J. F. Angulo, R. Devoret, and J. P. Claverys, Nucleic Acids Res. 20:6412, 1992). Exp10 is likely to be a membrane-associated protein, as it has a prokaryotic signal sequence and an Exp10-PhoA fusion localized with cell membranes. On the basis of sequence similarity, pneumococcal RecA is a member of bacterial RecA proteins responsible for homologous recombination of DNA. DNA-RNA hybridization analysis showed that this locus is transcribed as a polycistronic message, with increased transcription occurring during competence. With an Exp10-PhoA chimera used as a reporter, there was a 10-fold increase in the expression of the rec locus during competence while there was only minimal expression under growth conditions that repressed competence. The TFN- mutant containing the exp10-phoA fusion produced activator, a small extracellular polypeptide that induces competence, and the expression of rec was induced in response to activator. Therefore, the rec locus is directly required for genetic transformation and is regulated by the cell signaling mechanism that induces competence.
Pearce
B J
BJ
Laboratory of Molecular Infectious Diseases, Rockefeller University, New York, New York 10021.
Naughton
A M
AM
Campbell
E A
EA
Masure
H R
HR
D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000235:genetics; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D015964:Gene Expression Regulation, Bacterial; D008565:Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D009876:Operon / Q000235:genetics*; D011938:Rec A Recombinases / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011995:Recombination, Genetic / Q000235:genetics; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D013296:Streptococcus pneumoniae / Q000235:genetics*; D014158:Transcription, Genetic; D014170:Transformation, Genetic / Q000235:genetics*
D001426:Bacterial Proteins; C091161:CinA protein, Streptococcus pneumoniae; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D011938:Rec A Recombinases; D000469:Alkaline Phosphatase