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dc.contributor.authorHoskins, Aaron A.
dc.contributor.authorRodgers, Margaret L.
dc.contributor.authorFriedman, Larry J.
dc.contributor.authorGelles, Jeff
dc.contributor.authorMoore, Melissa J.
dc.date2022-08-11T08:09:45.000
dc.date.accessioned2022-08-23T16:42:07Z
dc.date.available2022-08-23T16:42:07Z
dc.date.issued2016-05-31
dc.date.submitted2016-08-16
dc.identifier.citation<p>Elife. 2016 May 31;5. pii: e14166. doi: 10.7554/eLife.14166. <a href="http://dx.doi.org/10.7554/eLife.14166">Link to article on publisher's site</a></p>
dc.identifier.issn2050-084X (Linking)
dc.identifier.doi10.7554/eLife.14166
dc.identifier.pmid27244240
dc.identifier.urihttp://hdl.handle.net/20.500.14038/40024
dc.description.abstractThe spliceosome is a complex machine composed of small nuclear ribonucleoproteins (snRNPs) and accessory proteins that excises introns from pre-mRNAs. After assembly the spliceosome is activated for catalysis by rearrangement of subunits to form an active site. How this rearrangement is coordinated is not well-understood. During activation, U4 must be released to allow U6 conformational change, while Prp19 complex (NTC) recruitment is essential for stabilizing the active site. We used multi-wavelength colocalization single molecule spectroscopy to directly observe the key events in Saccharomyces cerevisiae spliceosome activation. Following binding of the U4/U6.U5 tri-snRNP, the spliceosome either reverses assembly by discarding tri-snRNP or proceeds to activation by irreversible U4 loss. The major pathway for NTC recruitment occurs after U4 release. ATP stimulates both the competing U4 release and tri-snRNP discard processes. The data reveal the activation mechanism and show that overall splicing efficiency may be maintained through repeated rounds of disassembly and tri-snRNP reassociation.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=27244240&dopt=Abstract">Link to Article in PubMed</a></p>
dc.rightsCopyright © 2016, Hoskins et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectRNA
dc.subjectS. cerevisiae
dc.subjectbiochemistry
dc.subjectbiophysics
dc.subjectfluorescence
dc.subjectsingle-molecule
dc.subjectsnRNP
dc.subjectspliceosome
dc.subjectsplicing
dc.subjectstructural biology
dc.subjectBiochemistry
dc.subjectBiophysics
dc.subjectStructural Biology
dc.titleSingle molecule analysis reveals reversible and irreversible steps during spliceosome activation
dc.typeJournal Article
dc.source.journaltitleeLife
dc.source.volume5
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=3835&amp;context=oapubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/2830
dc.identifier.contextkey8985347
refterms.dateFOA2022-08-23T16:42:07Z
html.description.abstract<p>The spliceosome is a complex machine composed of small nuclear ribonucleoproteins (snRNPs) and accessory proteins that excises introns from pre-mRNAs. After assembly the spliceosome is activated for catalysis by rearrangement of subunits to form an active site. How this rearrangement is coordinated is not well-understood. During activation, U4 must be released to allow U6 conformational change, while Prp19 complex (NTC) recruitment is essential for stabilizing the active site. We used multi-wavelength colocalization single molecule spectroscopy to directly observe the key events in Saccharomyces cerevisiae spliceosome activation. Following binding of the U4/U6.U5 tri-snRNP, the spliceosome either reverses assembly by discarding tri-snRNP or proceeds to activation by irreversible U4 loss. The major pathway for NTC recruitment occurs after U4 release. ATP stimulates both the competing U4 release and tri-snRNP discard processes. The data reveal the activation mechanism and show that overall splicing efficiency may be maintained through repeated rounds of disassembly and tri-snRNP reassociation.</p>
dc.identifier.submissionpathoapubs/2830
dc.contributor.departmentDepartment of Biochemistry and Molecular Pharmacology


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Copyright © 2016, Hoskins et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
Except where otherwise noted, this item's license is described as Copyright © 2016, Hoskins et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.