High-resolution phenotypic landscape of the RNA Polymerase II trigger loop
High-resolution phenotypic landscape of the RNA Polymerase II trigger loop
The active site of multicellular 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 major 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.
Erinne Olivia C.、Jin Huiyan、Vandeventer Paul J.、Strohner Ralf、Van den Brulle Jan、Babu Sabareesh Ganesh、Qiu Chenxi、Lam Kenny C.、Cui Ping、Kaplan Craig D.、Dave Jui、Tang Leung K.、Muthukrishnan Nandhini、Sze Sing-Hoi
Mississippi State UniversityDepartment of Biochemistry & Biophysics, Texas A&M University||Current address: Roche NimblegenDepartment of Biochemistry & Biophysics, Texas A&M University||Current address: Department of Biochemistry & Molecular Biology, Baylor College of MedicineMorphoSys AG, PlaneggMorphoSys AG, PlaneggDepartment of Biochemistry & Biophysics, Texas A&M UniversityDepartment of Biochemistry & Biophysics, Texas A&M UniversityDepartment of Biochemistry & Biophysics, Texas A&M University||Current address: College of Medicine, Texas A&M Health Science CenterDepartment of Biochemistry & Biophysics, Texas A&M University||Current address: Department of Veterinary Pathobiology, Texas A&M UniversityDepartment of Biochemistry & Biophysics, Texas A&M UniversityDepartment of Biochemistry & Biophysics, Texas A&M University||Current address: Yale Cardiovascular Research CenterDepartment of Biochemistry & Biophysics, Texas A&M University||Current address: Department of Veterinary Pathobiology, Texas A&M UniversityDepartment of Biochemistry & Biophysics, Texas A&M University||Current address: Shire PharmaceuticalDepartment of Biochemistry & Biophysics, Texas A&M University||Department of Computer Science and Engineering, Texas A&M University
分子生物学生物化学生物物理学
Erinne Olivia C.,Jin Huiyan,Vandeventer Paul J.,Strohner Ralf,Van den Brulle Jan,Babu Sabareesh Ganesh,Qiu Chenxi,Lam Kenny C.,Cui Ping,Kaplan Craig D.,Dave Jui,Tang Leung K.,Muthukrishnan Nandhini,Sze Sing-Hoi.High-resolution phenotypic landscape of the RNA Polymerase II trigger loop[EB/OL].(2025-03-28)[2025-04-30].https://www.biorxiv.org/content/10.1101/068726.点此复制
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