Live-cell single-molecule dynamics of eukaryotic RNA polymerase machineries

• 成果类型: Article
• 署名作者: Ling, Yick Hin; Liang, Chloe; Wang, Sixiang; Wu, Carl
• 署名单位: Johns Hopkins University; Johns Hopkins University; Johns Hopkins Medicine
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-8075; 1095-9203
• DOI: 10.1126/science.ads0960
• 发表日期: 2026-03-19
• 页码: eads0960
• 关键词: CARBOXYL-TERMINAL DOMAIN; TRANSCRIPTION FACTOR TFIIB; TATA-BINDING PROTEIN; II CTD; saccharomyces-cerevisiae; in-vitro; complex; elongation; initiation; methylation

摘要

Eukaryotic gene expression is orchestrated by RNA polymerases (RNAPI, II, and III) and associated factors, yet their real-time dynamics remain obscure. Using single-molecule tracking in living yeast, we quantified the kinetics of 58 proteins encompassing three RNAP machineries. RNAPI and RNAPIII preinitiation complexes (PICs) engage in long-lived chromatin interactions, contrasting with transient RNAPII PICs. We further report kinetics of RNAPII-associated factors for elongation, histone modification, C-terminal domain (CTD) modification, RNA processing, and termination. Many elongation factors show brief rather than persistent association, suggesting dynamic interactions with factor exchange and allowing a potential repertoire of regulatory events. CTD truncation reduces U1 small nuclear ribonucleoprotein residence time and intron retention in ribosomal protein genes, providing insights into cotranscriptional splicing. Our findings establish a framework of dynamic interactions of RNAP machineries.