Identification and regulation of cold-inducible factors of Bordetella bronchiseptica.
The expression of bacterial cold-shock proteins (CSPs) is highly induced in response to cold shock, and some CSPs are essential for cells to resume growth at low temperature. Bordetella bronchiseptica encodes five CSPs (named CspA to CspE) with significant amino acid homology to CspA of Escherichia coli. In contrast to E. coli, the insertional knock-out of a single csp gene (cspB) strongly affected growth of B. bronchiseptica independent of temperature. In the case of three of the csp genes (cspA, cspB, cspC) more than one specific transcript could be detected. The net amount of cspA, cspB and cspC transcripts increased strongly after cold shock, while no such effect could be observed for cspD and cspE. The exposure to other stress conditions, including translation inhibitors, heat shock, osmotic stress and nutrient deprivation in the stationary phase, indicated that the csp genes are also responsive to these conditions. The coding regions of all of the cold-shock genes are preceded by a long non-translated upstream region (5'-UTR). In the case of the cspB gene, a deletion of parts of this region led to a significant reduction of translation of the resulting truncated transcript, indicating a role of the 5'-UTR in translational control. The cold-shock stimulon was investigated by 2D-PAGE and mass spectrometric characterization, leading to the identification of additional cold-inducible proteins (CIPs). Interestingly, two cold-shock genes (cspC and cspD) were found to be under the negative control of the BvgAS system, the main transcriptional regulator of Bordetella virulence genes. Moreover, a negative effect of slight overexpression of CspB, but not of the other CSPs, on the transcription of the adenylate cyclase toxin CyaA of Bordetella pertussis was observed, suggesting cross-talk between the CSP-mediated stress response stimulon and the Bordetella virulence regulon.
Stübs
Dorothee
D
Lehrstuhl für Mikrobiologie, Biozentrum der Universität Würzburg, Am Hubland, D-97074 Würzburg, Germany.
Fuchs
Thilo M
TM
Zentralinstitut für Ernährungs- und Lebensmittelforschung, Abteilung Mikrobiologie, D-85354 Freising, Germany.
Schneider
Boris
B
Lehrstuhl für Mikrobiologie, Biozentrum der Universität Würzburg, Am Hubland, D-97074 Würzburg, Germany.
Bosserhoff
Armin
A
Zentrum für Molekulare Biologie, Universität Heidelberg, D-69120 Heidelberg, Germany.
Gross
Roy
R
Lehrstuhl für Mikrobiologie, Biozentrum der Universität Würzburg, Am Hubland, D-97074 Würzburg, Germany.
D020121:5' Untranslated Regions / Q000235:genetics; D000222:Adaptation, Physiological; D037361:Adenylate Cyclase Toxin / Q000096:biosynthesis; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D016950:Bordetella bronchiseptica / Q000235:genetics / Q000502:physiology*; D003080:Cold Temperature; D015180:Electrophoresis, Gel, Two-Dimensional; D015964:Gene Expression Regulation, Bacterial; D006360:Heat-Shock Proteins / Q000235:genetics / Q000502:physiology; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D020543:Proteome / Q000032:analysis; D012329:RNA, Bacterial / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D018085:Regulon; D017384:Sequence Deletion / Q000502:physiology; D017385:Sequence Homology; D014157:Transcription Factors / Q000502:physiology; D014158:Transcription, Genetic
D020121:5' Untranslated Regions; D037361:Adenylate Cyclase Toxin; D001426:Bacterial Proteins; C065601:BvgA protein, Bacteria; C088099:CspC protein, bacteria; C088098:CspD protein, bacteria; D006360:Heat-Shock Proteins; D020543:Proteome; D012329:RNA, Bacterial; D012333:RNA, Messenger; D014157:Transcription Factors; C080662:bvgS protein, Bordetella pertussis; C062260:cold shock protein CS7.4, Bacteria; C077188:cold-shock protein CspB, Bacteria