Ochs, Simon M. and Thumann, Sybille and Richau, Renate and Weirauch, Matt T. and Lowe, Todd M. and Thomm, Michael and Hausner, Winfried (2012) Activation of Archaeal Transcription Mediated by Recruitment of Transcription Factor B. JOURNAL OF BIOLOGICAL CHEMISTRY, 287 (22). pp. 18863-18871. ISSN 0021-9258,
Full text not available from this repository. (Request a copy)Abstract
Archaeal promoters consist of a TATA box and a purine-rich adjacent upstream sequence (transcription factor B (TFB)-responsive element (BRE)), which are bound by the transcription factors TATA box-binding protein (TBP) and TFB. Currently, only a few activators of archaeal transcription have been experimentally characterized. The best studied activator, Ptr2, mediates activation by recruitment of TBP. Here, we present a detailed biochemical analysis of an archaeal transcriptional activator, PF1088, which was identified in Pyrococcus furiosus by a bioinformatic approach. Operon predictions suggested that an upstream gene, pf1089, is polycistronically transcribed with pf1088. We demonstrate that PF1088 stimulates in vitro transcription by up to 7-fold when the pf1089 promoter is used as a template. By DNase I and hydroxyl radical footprinting experiments, we show that the binding site of PF1088 is located directly upstream of the BRE of pf1089. Mutational analysis indicated that activation requires the presence of the binding site for PF1088. Furthermore, we show that activation of transcription by PF1088 is dependent upon the presence of an imperfect BRE and is abolished when the pf1089 BRE is replaced with a BRE from a strong archaeal promoter. Gel shift experiments showed that TFB recruitment to the pf1089 operon is stimulated by PF1088, and TFB seems to stabilize PF1088 operator binding even in the absence of TBP. Taken together, these results represent the first biochemical evidence for a transcriptional activator working as a TFB recruitment factor in Archaea, for which the designation TFB-RF1 is suggested.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CELL-FREE TRANSCRIPTION; TATA-BINDING PROTEIN; RNA-POLYMERASE; PYROCOCCUS-FURIOSUS; SULFOLOBUS-SOLFATARICUS; PREINITIATION COMPLEX; INITIATION COMPLEX; STRUCTURAL BASIS; TFIIE-ALPHA; DNA-BINDING; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 13 May 2020 09:51 |
| Last Modified: | 13 May 2020 09:51 |
| URI: | https://pred.uni-regensburg.de/id/eprint/18715 |
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