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Journal of Bacteriology, May 2005, p. 3339-3351, Vol. 187, No. 10
0021-9193/05/$08.00+0     doi:10.1128/JB.187.10.3339-3351.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Global Transcriptional and Proteomic Analysis of the Sig1 Heat Shock Regulon of Deinococcus radiodurans{dagger}

Amy K. Schmid,1* Heather A. Howell,2 John R. Battista,3 Scott N. Peterson,2,4 and Mary E. Lidstrom1,5,6

Program in Molecular and Cellular Biology, University of Washington, Seattle, Washington 98195-2180,1 The Institute for Genomic Research, 9712 Medical Center Dr., Rockville, Maryland 20850,2 Department of Biological Sciences, Louisiana State University and A&M College, Baton Rouge, Louisiana 70803,3 Department of Biochemistry and Molecular Biology, George Washington University, Washington, D.C. 20052,4 Department of Chemical Engineering,5 Department of Microbiology, University of Washington, Seattle, Washington6

Received 18 October 2004/ Accepted 4 February 2005

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 {sigma}70-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.

* Corresponding author. Current address: Institute for Systems Biology, 1441 34th St., Seattle, WA 98103. Phone: (206) 732-1493. Fax: (206) 732-1299. E-mail: aschmid{at}systemsbiology.org.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, May 2005, p. 3339-3351, Vol. 187, No. 10
0021-9193/05/$08.00+0     doi:10.1128/JB.187.10.3339-3351.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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