CovS/CovR of group B streptococcus: a two-component global regulatory system involved in virulence.
In this study, we carried out a detailed structural and functional analysis of a Streptococcus agalactiae (GBS) two-component system which is orthologous to the CovS/CovR (CsrS/CsrR) regulatory system of Streptococcus pyogenes. In GBS, covR and covS are part of a seven gene operon transcribed from two promoters that are not regulated by CovR. A DeltacovSR mutant was found to display dramatic phenotypic changes such as increased haemolytic activity and reduced CAMP activity on blood agar. Adherence of the DeltacovSR mutant to epithelial cells was greatly increased and analysis by transmission electron microscopy revealed the presence at its surface of a fibrous extracellular matrix that might be involved in these intercellular interactions. However, the DeltacovSR mutant was unable to initiate growth in RPMI and its viability in human normal serum was greatly impaired. A major finding of this phenotypic analysis was that the CovS/CovR system is important for GBS virulence, as a 3 log increase of the LD(50) of the mutant strain was observed in the neonate rat sepsis model. The pleiotropic phenotype of the DeltacovSR mutant is in full agreement with the large number of genes controlled by CovS/CovR as seen by expression profiling analysis, many of which encode potentially secreted or cell surface-associated proteins: 76 genes are repressed whereas 63 were positively regulated. CovR was shown to bind directly to the regulatory regions of several of these genes and a consensus CovR recognition sequence was proposed using both DNase I footprinting and computational analyses.
Lamy
Marie-Cécile
MC
INSERM U-570, Faculté de Médecine Necker-Enfants Malades, 75730 Paris Cedex 15, France.
Zouine
Mohammed
M
Fert
Juliette
J
Vergassola
Massimo
M
Couve
Elisabeth
E
Pellegrini
Elisabeth
E
Glaser
Philippe
P
Kunst
Frank
F
Msadek
Tarek
T
Trieu-Cuot
Patrick
P
Poyart
Claire
C
D000818:Animals; D001422:Bacterial Adhesion / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000382:microbiology; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D006460:Hemolysin Proteins; D006461:Hemolysis; D006801:Humans; D007928:Lethal Dose 50; D009876:Operon; D011401:Promoter Regions, Genetic; D011494:Protein Kinases / Q000235:genetics / Q000502:physiology*; D051381:Rats; D018085:Regulon; D012097:Repressor Proteins / Q000235:genetics / Q000502:physiology*; D015398:Signal Transduction / Q000235:genetics; D013290:Streptococcal Infections / Q000382:microbiology; D013292:Streptococcus agalactiae / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity* / Q000648:ultrastructure; D014158:Transcription, Genetic; D014774:Virulence / Q000235:genetics*; D037521:Virulence Factors / Q000235:genetics*
D001426:Bacterial Proteins; C039092:CAMP protein, Streptococcus; C000619936:CovR protein, Streptococcus agalactiae; D006460:Hemolysin Proteins; D012097:Repressor Proteins; D037521:Virulence Factors; D011494:Protein Kinases; C432144:CsrS protein, bacteria