Keese, Annette M. and Schut, Gerrit J. and Ouhammouch, Mohamed and Adams, Michael W. W. and Thomm, Michael (2010) Genome-Wide Identification of Targets for the Archaeal Heat Shock Regulator Phr by Cell-Free Transcription of Genomic DNA. JOURNAL OF BACTERIOLOGY, 192 (5). pp. 1292-1298. ISSN 0021-9193, 1098-5530
Full text not available from this repository. (Request a copy)Abstract
The hyperthermophilic archaeon Pyrococcus furiosus grows optimally near 100 degrees C and undergoes a heat shock response at 105 degrees C, mediated at least in part by the heat shock regulator Phr. Genes encoding a small heat shock protein (HSP20) and a member of the AAA(+) ATPase are the only known targets of the regulator, but a genetic mutant of Phr has yet to be characterized. We describe here an alternative approach for the identification of the regulon of Phr based on cell-free transcription of fragmented chromosomal DNA in the presence or absence of the regulator and hybridization of in vitro RNA to P. furiosus whole-genome microarrays. Our results confirmed the phr, the hsp20, and the aaa(+) ATPase genes as targets of Phr and also identified six additional open reading frames, PF0624, PF1042, PF1291, PF1292, PF1488, and PF1616, as Phr-responsive genes, which include that encoding di-myo-inositol phosphate synthase. Transcription of the identified novel genes was inhibited by Phr in standard transcription assays, and the novel consensus sequence 5'-TTTAnnn ACnnnnnGTnAnnAAAA-3' (uppercase letters denote a high conservation of the bases) was inferred from our data as the Phr recognition motif. Mutational evidence for the significance of this sequence as Phr recognition was provided in DNA-binding experiments.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PYROCOCCUS-FURIOSUS; HYPERTHERMOPHILIC ARCHAEA; IN-VITRO; PROTEIN; CHAPERONIN; REGULON; ROMA; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) > Prof. Dr. Michael Thomm |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 05 Aug 2020 07:37 |
| Last Modified: | 05 Aug 2020 07:37 |
| URI: | https://pred.uni-regensburg.de/id/eprint/25082 |
Actions (login required)
 |
View Item |
