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High-Resolution Phenotypic Landscape of the RNA Polymerase II Trigger Loop.
- Source :
-
PLoS genetics [PLoS Genet] 2016 Nov 29; Vol. 12 (11), pp. e1006321. Date of Electronic Publication: 2016 Nov 29 (Print Publication: 2016). - Publication Year :
- 2016
-
Abstract
- The active sites of multisubunit RNA polymerases have a "trigger loop" (TL) that multitasks in substrate selection, catalysis, and translocation. To dissect the Saccharomyces cerevisiae RNA polymerase II TL at individual-residue resolution, we quantitatively phenotyped nearly all TL single variants en masse. Three mutant classes, revealed by phenotypes linked to transcription defects or various stresses, have distinct distributions among TL residues. We find that mutations disrupting an intra-TL hydrophobic pocket, proposed to provide a mechanism for substrate-triggered TL folding through destabilization of a catalytically inactive TL state, confer phenotypes consistent with pocket disruption and increased catalysis. Furthermore, allele-specific genetic interactions among TL and TL-proximal domain residues support the contribution of the funnel and bridge helices (BH) to TL dynamics. Our structural genetics approach incorporates structural and phenotypic data for high-resolution dissection of transcription mechanisms and their evolution, and is readily applicable to other essential yeast proteins.<br />Competing Interests: Authors JVdB and RS are employed by MorphoSys AG, which currently holds rights to the Slonomics DNA synthesis technology utilized in the manuscript.
- Subjects :
- Alleles
Catalysis
Catalytic Domain genetics
Crystallography, X-Ray
Mutant Proteins chemistry
Mutation
Protein Folding
Protein Structure, Secondary
Protein Transport genetics
RNA Polymerase II chemistry
Saccharomyces cerevisiae enzymology
Substrate Specificity
Mutant Proteins genetics
RNA Polymerase II genetics
Saccharomyces cerevisiae genetics
Transcription, Genetic
Subjects
Details
- Language :
- English
- ISSN :
- 1553-7404
- Volume :
- 12
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- PLoS genetics
- Publication Type :
- Academic Journal
- Accession number :
- 27898685
- Full Text :
- https://doi.org/10.1371/journal.pgen.1006321