Reich, Christoph and Zeller, Mirijam and Milkereit, Philipp and Hausner, Winfried and Cramer, Patrick and Tschochner, Herbert and Thomm, Michael (2009) The archaeal RNA polymerase subunit P and the eukaryotic polymerase subunit Rpb12 are interchangeable in vivo and in vitro. MOLECULAR MICROBIOLOGY, 71 (4). pp. 989-1002. ISSN 0950-382X, 1365-2958
Full text not available from this repository. (Request a copy)Abstract
The general subunit of all three eukaryotic RNA polymerases, Rpb12, and subunit P of the archaeal enzyme show sequence similarities in their N-terminal zinc ribbon and some highly conserved residues in the C-terminus. We report here that archaeal subunit P under the control of a strong yeast promoter could complement the lethal phenotype of a RPB12 deletion mutant and that subunit Rpb12 from yeast can functionally replace subunit P during reconstitution of the archaeal RNA polymerase. The Delta P enzyme is unable to form stable open complexes, but can efficiently extend a dinucleotide on a premelted template or RNA on an elongation scaffold. This suggests that subunit P is directly or indirectly involved in promoter opening. The activity of the Delta P enzyme can be rescued by the addition of Rpb12 or subunit P to transcription reactions. Mutation of cysteine residues in the zinc ribbon impair the activity of the enzyme in several assays and this mutated form of P is rapidly replaced by wild-type P in transcription reactions. The conserved zinc ribbon in the N-terminus seems to be important for proper interaction of the complete subunit with other RNA polymerase subunits and a 17-amino-acid C-terminal peptide is sufficient to support all basic RNA polymerase functions in vitro.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | TRANSCRIPTION PREINITIATION COMPLEX; SACCHAROMYCES-CEREVISIAE; SHUTTLE VECTORS; YEAST; DNA; ELONGATION; ABC10-ALPHA; MACHINERY; MECHANISM; PROTEINS; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III > Prof. Dr. Herbert Tschochner Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III > Dr. Philipp Milkereit Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) > Prof. Dr. Michael Thomm |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 07 Oct 2020 06:33 |
| Last Modified: | 07 Oct 2020 06:33 |
| URI: | https://pred.uni-regensburg.de/id/eprint/29538 |
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