Gerber, Jochen and Reiter, Alarich and Steinbauer, Robert and Jakob, Steffen and Kuhn, Claus-Dieter and Cramer, Patrick and Griesenbeck, Joachim and Milkereit, Philipp and Tschochner, Herbert (2008) Site specific phosphorylation of yeast RNA polymerase I. NUCLEIC ACIDS RESEARCH, 36 (3). pp. 793-802. ISSN 0305-1048, 1362-4962
Full text not available from this repository. (Request a copy)Abstract
All nuclear RNA polymerases are phosphoprotein complexes. Yeast RNA polymerase I (Pol I) contains approximately 15 phosphate groups, distributed to 5 of the 14 subunits. Information about the function of the single phosphosites and their position in the primary, secondary and tertiary structure is lacking. We used a rapid and efficient way to purify yeast RNA Pol I to determine 13 phosphoserines and threonines. Seven of these phosphoresidues could be located in the 3D-homology model for Pol I, five of them are more at the surface. The single phosphorylated residues were systematically mutated and the resulting strains and Pol I preparations were analyzed in cellular growth, Pol I composition, stability and genetic interaction with non-essential components of the transcription machinery. Surprisingly, all Pol I phosphorylations analyzed were found to be non-essential post-translational modifications. However, one mutation (subunit A190 S685D) led to higher growth rates in the presence of 6AU or under environmental stress conditions, and was synthetically lethal with a deletion of the Pol I subunit A12.2, suggesting a role in RNA cleavage/elongation or termination. Our results suggest that individual major or constitutively phosphorylated residues contribute to non-essential Pol I-functions.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | SACCHAROMYCES-CEREVISIAE ENCODES; MASS-SPECTROMETRY; STRUCTURAL BASIS; SUBUNIT COMMON; ACTIVE-CENTER; TRANSCRIPTION; GENE; SUBCOMPLEX; PROTEIN; PHOSPHOPROTEOME; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III > Prof. Dr. Herbert Tschochner |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 10 Nov 2020 07:52 |
| Last Modified: | 10 Nov 2020 07:52 |
| URI: | https://pred.uni-regensburg.de/id/eprint/31446 |
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