Rpb9 subunit controls transcription fidelity by delaying NTP sequestration in RNA polymerase II.
Rpb9 is a small non-essential subunit of yeast RNA polymerase II located on the surface on the enzyme. Deletion of the RPB9 gene shows synthetic lethality with the low fidelity rpb1-E1103G mutation localized in the trigger loop, a mobile element of the catalytic Rpb1 subunit, which has been shown to ... control transcription fidelity. Similar to the rpb1-E1103G mutation, the RPB9 deletion substantially enhances NTP misincorporation and increases the rate of mismatch extension with the next cognate NTP in vitro. Using pre-steady state kinetic analysis, we show that RPB9 deletion promotes sequestration of NTPs in the polymerase active center just prior to the phosphodiester bond formation. We propose a model in which the Rpb9 subunit controls transcription fidelity by delaying the closure of the trigger loop on the incoming NTP via interaction between the C-terminal domain of Rpb9 and the trigger loop. Our findings reveal a mechanism for regulation of transcription fidelity by protein factors located at a large distance from the active center of RNA polymerase II.
Mesh Terms:
Adenosine Triphosphate, Base Sequence, Cytidine Triphosphate, Fungal Proteins, Kinetics, Models, Molecular, Mutation, Nucleotides, Protein Conformation, Protein Subunits, RNA Polymerase II, Time Factors, Transcription, Genetic, Uridine Triphosphate
Adenosine Triphosphate, Base Sequence, Cytidine Triphosphate, Fungal Proteins, Kinetics, Models, Molecular, Mutation, Nucleotides, Protein Conformation, Protein Subunits, RNA Polymerase II, Time Factors, Transcription, Genetic, Uridine Triphosphate
J. Biol. Chem.
Date: Jul. 17, 2009
PubMed ID: 19439405
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