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首页|Multi-Factor Authentication of Potential 5’ Splice Sites by the Saccharomyces cerevisiae U1 snRNP

Multi-Factor Authentication of Potential 5’ Splice Sites by the Saccharomyces cerevisiae U1 snRNP

Multi-Factor Authentication of Potential 5’ Splice Sites by the Saccharomyces cerevisiae U1 snRNP

来源:bioRxiv_logobioRxiv
英文摘要

ABSTRACT In eukaryotes, splice sites define the introns of pre-mRNAs and must be recognized and excised with nucleotide precision by the spliceosome to make the correct mRNA product. In one of the earliest steps of spliceosome assembly, the U1 small nuclear ribonucleoprotein (snRNP) recognizes the 5’ splice site (5’ SS) through a combination of base pairing, protein-RNA contacts, and interactions with other splicing factors. Previous studies investigating the mechanisms of 5’ SS recognition have largely been done in vivo or in cellular extracts where the U1/5’ SS interaction is difficult to deconvolute from the effects of trans-acting factors or RNA structure. In this work we used co-localization single-molecule spectroscopy (CoSMoS) to elucidate the pathway of 5’ SS selection by purified yeast U1 snRNP. We determined that U1 reversibly selects 5’ SS in a sequence-dependent, two-step mechanism. A kinetic selection scheme enforces pairing at particular positions rather than overall duplex stability to achieve long-lived U1 binding. Our results provide a kinetic basis for how U1 may rapidly surveil nascent transcripts for 5’ SS and preferentially accumulate at these sequences rather than on close cognates. IMPACT STATEMENTThe yeast U1 snRNP recognizes multiple features of target RNAs to reversibly identify splicing-competent 5’ splice sites.

Hansen Sarah R.、Corr¨oa Ivan R. Jr、Hoskins Aaron A.、Smith Lloyd M.、Scalf Mark

Department of Biochemistry||Integrated Program in BiochemistryNew England BiolabsDepartment of Biochemistry||Integrated Program in Biochemistry||Department of ChemistryDepartment of ChemistryDepartment of Chemistry

10.1101/2021.05.18.443434

分子生物学细胞生物学生物科学研究方法、生物科学研究技术

SplicingSingle Molecule FluorescenceCoSMoSSpliceosomesnRNPRNA

Hansen Sarah R.,Corr¨oa Ivan R. Jr,Hoskins Aaron A.,Smith Lloyd M.,Scalf Mark.Multi-Factor Authentication of Potential 5’ Splice Sites by the Saccharomyces cerevisiae U1 snRNP[EB/OL].(2025-03-28)[2025-05-02].https://www.biorxiv.org/content/10.1101/2021.05.18.443434.点此复制

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