The group A streptococcal virR49 gene controls expression of four structural vir regulon genes.
Within a genomic locus termed the vir regulon, virR genes of opacity factor-nonproducing (OF-) group A streptococci (GAS) are known to control the expression of the genes encoding M protein (emm) and C5a peptidase (scpA) and of virR itself. Within the corresponding genomic locus, opacity factor-producing (OF+) GAS harbor additional emm-related genes encoding immunoglobulin G- and immunoglobulin A-binding proteins (fcrA and enn, respectively). The virR gene region of the OF+ GAS M-type 49 strain CS101 was amplified by PCR, and 2,650 bp were directly sequenced. An open reading frame of 1,599 bp exhibited 76% overall homology to published virR sequences. By utilizing mRNA analysis, the 5' ends of two specific transcripts were mapped 370 and 174 bp upstream of the start codon of this open reading frame. The deduced sequences of the corresponding promoters and their locations differed from those of previously reported virR promoters. Transcripts from wild-type fcrA49, emm49, enn49, and scpA49 genes located downstream of virR49 were characterized as being monocistronic. The transcripts were quantified and mapped for their 5' ends. Subsequently, the virR49 gene was inactivated by specific insertion of a nonreplicative pSF152 vector containing recombinant virR49 sequences. The RNA from the resulting vir-mut strain did not contain transcripts of virR49, fcrA49, emm49, or enn49 and contained reduced amounts of the scpA49 transcript when compared with wild-type RNA. The mRNA control from the streptokinase gene was demonstrated not to be affected. When strain vir-mut was rotated in human blood, it was found to be fully sensitive to phagocytosis by human leukocytes. Thus, the present study provides evidence that virR genes in OF+ GAS could be involved in the control of up to five vir regulon genes, and their unaffected regulatory activity is associated with features postulated as crucial for GAS virulence.
Podbielski
A
A
Institute of Medical Microbiology, Technical University (RWTH), Aachen, Germany.
Flosdorff
A
A
Weber-Heynemann
J
J
D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D002874:Chromosome Mapping; D010450:Endopeptidases / Q000096:biosynthesis / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D018085:Regulon / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013297:Streptococcus pyogenes / Q000235:genetics* / Q000472:pathogenicity; D013300:Streptokinase / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic; D014774:Virulence / Q000235:genetics
D018829:Adhesins, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D012333:RNA, Messenger; C021908:opacity factor; C014689:streptococcal M protein; C068267:mry protein, Streptococcus pyogenes; D010450:Endopeptidases; D010447:Peptide Hydrolases; D013300:Streptokinase; C053088:C5a peptidase