Merkl, Philipp E. and Pilsl, Michael and Fremter, Tobias and Schwank, Katrin and Engel, Christoph and Laengst, Gernot and Milkereit, Philipp and Griesenbeck, Joachim and Tschochner, Herbert (2020) RNA polymerase I (Pol I) passage through nucleosomes depends on Pol I subunits binding its lobe structure. JOURNAL OF BIOLOGICAL CHEMISTRY, 295 (15). pp. 4782-4795. ISSN 0021-9258, 1083-351X
Full text not available from this repository. (Request a copy)Abstract
RNA polymerase I (Pol I) is a highly efficient enzyme specialized in synthesizing most ribosomal RNAs. After nucleosome deposition at each round of rDNA replication, the Pol I transcription machinery has to deal with nucleosomal barriers. It has been suggested that Pol I?associated factors facilitate chromatin transcription, but it is unknown whether Pol I has an intrinsic capacity to transcribe through nucleosomes. Here, we used in vitro transcription assays to study purified WT and mutant Pol I variants from the yeast Saccharomyces cerevisiae and compare their abilities to pass a nucleosomal barrier with those of yeast Pol II and Pol III. Under identical conditions, purified Pol I and Pol III, but not Pol II, could transcribe nucleosomal templates. Pol I mutants lacking either the heterodimeric subunit Rpa34.5/Rpa49 or the C-terminal part of the specific subunit Rpa12.2 showed a lower processivity on naked DNA templates, which was even more reduced in the presence of a nucleosome. Our findings suggest that the lobe-binding subunits Rpa34.5/Rpa49 and Rpa12.2 facilitate passage through nucleosomes, suggesting possible cooperation among these subunits. We discuss the contribution of Pol I?specific subunit domains to efficient Pol I passage through nucleosomes in the context of transcription rate and processivity.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | TRANSCRIPTION INITIATION; SACCHAROMYCES-CEREVISIAE; CHROMATIN TRANSCRIPTION; MOLECULAR-STRUCTURES; TOPOISOMERASE-I; ELONGATION RATE; CROSS-LINKING; PAF1 COMPLEX; ARCHITECTURE; TFIIF; nucleolus; RNA polymerase II; RNA polymerase I; ribosomal ribonucleic acid (rRNA) (ribosomal RNA); transcription; transcription elongation factor; RNA polymerase III; nucleosome; ribosome; chromatin; Rpa34; 5; Rpa49 |
| 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. Joachim Griesenbeck Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III > Dr. Philipp Milkereit |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 26 Mar 2021 12:40 |
| Last Modified: | 26 Mar 2021 12:40 |
| URI: | https://pred.uni-regensburg.de/id/eprint/44750 |
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