SpdR, a response regulator required for stationary-phase induction of Caulobacter crescentus cspD.
The cold shock protein (CSP) family includes small polypeptides that are induced upon temperature downshift and stationary phase. The genome of the alphaproteobacterium Caulobacter crescentus encodes four CSPs, with two being induced by cold shock and two at the onset of stationary phase. In order to identify the environmental signals and cell factors that are involved in cspD expression at stationary phase, we have analyzed cspD transcription during growth under several nutrient conditions. The results showed that expression of cspD was affected by the medium composition and was inversely proportional to the growth rate. The maximum levels of expression were decreased in a spoT mutant, indicating that ppGpp may be involved in the signalization for carbon starvation induction of cspD. A Tn5 mutant library was screened for mutants with reduced cspD expression, and 10 clones that showed at least a 50% reduction in expression were identified. Among these, a strain with a transposon insertion into a response regulator of a two-component system showed no induction of cspD at stationary phase. This protein (SpdR) was able to acquire a phosphate group from its cognate histidine kinase, and gel mobility shift assay and DNase I footprinting experiments showed that it binds to an inverted repeat sequence of the cspD regulatory region. A mutated SpdR with a substitution of the conserved aspartyl residue that is the probable phosphorylation site is unable to bind to the cspD regulatory region and to complement the spdR mutant phenotype.
da Silva
Carolina A P T
CA
Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
Balhesteros
Heloise
H
Mazzon
Ricardo R
RR
Marques
Marilis V
MV
D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000096:biosynthesis*; D016935:Caulobacter crescentus / Q000235:genetics / Q000502:physiology*; D003470:Culture Media / Q000737:chemistry; D018983:DNA Footprinting; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D006159:Guanosine Tetraphosphate / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000096:biosynthesis*; D016254:Mutagenesis, Insertional; D020125:Mutation, Missense; D011485:Protein Binding; D011755:Pyrophosphatases / Q000235:genetics / Q000378:metabolism; D050436:Regulatory Elements, Transcriptional; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; C088098:CspD protein, bacteria; D003470:Culture Media; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D006360:Heat-Shock Proteins; D014157:Transcription Factors; D006159:Guanosine Tetraphosphate; C027627:guanosine-3',5'-bis(diphosphate) 3'-pyrophosphatase; D011755:Pyrophosphatases