Global transcriptional and proteomic analysis of the Sig1 heat shock regulon of Deinococcus radiodurans.
The sig1 gene, predicted to encode an extracytoplasmic function-type heat shock sigma factor of Deinococcus radiodurans, has been shown to play a central role in the positive regulation of the heat shock operons groESL and dnaKJ. To determine if Sig1 is required for the regulation of additional heat shock genes, we monitored the global transcriptional and proteomic profiles of a D. radiodurans R1 sig1 mutant and wild-type cells in response to elevated temperature stress. Thirty-one gene products were identified that showed heat shock induction in the wild type but not in the sig1 mutant. Quantitative real-time PCR experiments verified the transcriptional requirement of Sig1 for the heat shock induction of the mRNA of five of these genes-dnaK, groES, DR1314, pspA, and hsp20. hsp20 appears to encode a new member of the small heat shock protein superfamily, DR1314 is predicted to encode a hypothetical protein with no recognizable orthologs, and pspA is predicted to encode a protein involved in maintenance of membrane integrity. Deletion mutation analysis demonstrated the importance in heat shock protection of hsp20 and DR1314. The promoters of dnaKJE, groESL, DR1314, pspA, and hsp20 were mapped and, combined with computer-based pattern searches of the upstream regions of the 26 other Sig1 regulon members, these results suggested that Sig1 might recognize both sigma70-type and sigma(W)-type promoter consensus sequences. These results expand the D. radiodurans Sig1 heat shock regulon to include 31 potential new members, including not only factors with cytoplasmic functions, such as groES and dnaK, but also those with extracytoplasmic functions, like pspA.
Schmid
Amy K
AK
Program in Molecular and Cellular Biology, University of Washington, Seattle, Washington 98195-2180, USA. aschmid@systemsbiology.org
Howell
Heather A
HA
Battista
John R
JR
Peterson
Scott N
SN
Lidstrom
Mary E
ME
D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D018835:Chaperonin 10 / Q000235:genetics; D016384:Consensus Sequence; D034301:Deinococcus / Q000737:chemistry / Q000235:genetics*; D015180:Electrophoresis, Gel, Two-Dimensional; D029968:Escherichia coli Proteins / Q000235:genetics; D050886:HSP20 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D006360:Heat-Shock Proteins / Q000235:genetics; D018869:Heat-Shock Response; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010750:Phosphoproteins / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D040901:Proteomics; D018085:Regulon / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012808:Sigma Factor / Q000032:analysis / Q000235:genetics*; D013696:Temperature; D014158:Transcription, Genetic / Q000502:physiology*
D001426:Bacterial Proteins; D018835:Chaperonin 10; D029968:Escherichia coli Proteins; D050886:HSP20 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D010750:Phosphoproteins; D012808:Sigma Factor; C069816:phage shock protein, Bacteria; C063842:dnaK protein, E coli