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Site specific phosphorylation of yeast RNA polymerase I


Gerber, Jochen ; Reiter, Alarich ; Steinbauer, Robert ; Jakob, Stefan ; Kuhn, Claus-D. ; Cramer, Patrick ; Griesenbeck, Joachim ; Milkereit, Philipp ; Tschochner, Herbert:
Site specific phosphorylation of yeast RNA polymerase I.
In: Nucleic Acids Research. Bd. 36 (2008) Heft 3 . - S. 793-802.
ISSN 1362-4962
DOI: https://doi.org/10.1093/nar/gkm1093


Link zum Volltext (externe URL): Volltext


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.

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Publikationsform: Artikel in einer Zeitschrift
Begutachteter Beitrag: Ja
Keywords: Amino Acid Sequence; Fungal Proteins/*chemistry/genetics/metabolism; Models, Molecular; Molecular Sequence Data; Mutation; Phenotype; Phosphoproteins/*chemistry/genetics; Phosphorylation; Phosphoserine/analysis; Phosphothreonine/analysis; RNA Polymerase I/*chemistry/genetics/metabolism; Yeasts/enzymology
Institutionen der Universität: Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften > Fachgruppe Chemie
Profilfelder > Advanced Fields > Molekulare Biowissenschaften
Forschungseinrichtungen > Forschungszentren > Bayreuther Zentrum für Molekulare Biowissenschaften - BZMB
Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften
Profilfelder > Advanced Fields
Forschungseinrichtungen > Forschungszentren
Titel an der UBT entstanden: Nein
Themengebiete aus DDC: 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften; Biologie
Eingestellt am: 10 Jan 2017 11:27
Letzte Änderung: 10 Jan 2017 11:27
URI: https://eref.uni-bayreuth.de/id/eprint/35643