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
J. Biol. Chem.
Date: Jul. 17, 2009
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