Synergistic assembly of human pre-spliceosomes across introns and exons
Most human genes contain multiple introns, necessitating mechanisms to effectively define exons and ensure their proper connection by spliceosomes. Human spliceosome assembly involves both cross-intron and cross-exon interactions, but how these work together is unclear. We examined in human nuclear...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2018-06-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/37751 |
| _version_ | 1811201337027723264 |
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| author | Joerg E Braun Larry J Friedman Jeff Gelles Melissa J Moore |
| author_facet | Joerg E Braun Larry J Friedman Jeff Gelles Melissa J Moore |
| author_sort | Joerg E Braun |
| collection | DOAJ |
| description | Most human genes contain multiple introns, necessitating mechanisms to effectively define exons and ensure their proper connection by spliceosomes. Human spliceosome assembly involves both cross-intron and cross-exon interactions, but how these work together is unclear. We examined in human nuclear extracts dynamic interactions of single pre-mRNA molecules with individual fluorescently tagged spliceosomal subcomplexes to investigate how cross-intron and cross-exon processes jointly promote pre-spliceosome assembly. U1 subcomplex bound to the 5' splice site of an intron acts jointly with U1 bound to the 5' splice site of the next intron to dramatically increase the rate and efficiency by which U2 subcomplex is recruited to the branch site/3' splice site of the upstream intron. The flanking 5' splice sites have greater than additive effects implying distinct mechanisms facilitating U2 recruitment. This synergy of 5' splice sites across introns and exons is likely important in promoting correct and efficient splicing of multi-intron pre-mRNAs. |
| first_indexed | 2024-04-12T02:19:05Z |
| format | Article |
| id | doaj.art-87a46f33740a469c9492fc4e66136c1a |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:19:05Z |
| publishDate | 2018-06-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-87a46f33740a469c9492fc4e66136c1a2022-12-22T03:52:10ZengeLife Sciences Publications LtdeLife2050-084X2018-06-01710.7554/eLife.37751Synergistic assembly of human pre-spliceosomes across introns and exonsJoerg E Braun0https://orcid.org/0000-0002-8309-6401Larry J Friedman1https://orcid.org/0000-0003-4946-8731Jeff Gelles2https://orcid.org/0000-0001-7910-3421Melissa J Moore3RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United States; Department of Biochemistry, Brandeis University, Waltham, United StatesDepartment of Biochemistry, Brandeis University, Waltham, United StatesDepartment of Biochemistry, Brandeis University, Waltham, United StatesRNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, United StatesMost human genes contain multiple introns, necessitating mechanisms to effectively define exons and ensure their proper connection by spliceosomes. Human spliceosome assembly involves both cross-intron and cross-exon interactions, but how these work together is unclear. We examined in human nuclear extracts dynamic interactions of single pre-mRNA molecules with individual fluorescently tagged spliceosomal subcomplexes to investigate how cross-intron and cross-exon processes jointly promote pre-spliceosome assembly. U1 subcomplex bound to the 5' splice site of an intron acts jointly with U1 bound to the 5' splice site of the next intron to dramatically increase the rate and efficiency by which U2 subcomplex is recruited to the branch site/3' splice site of the upstream intron. The flanking 5' splice sites have greater than additive effects implying distinct mechanisms facilitating U2 recruitment. This synergy of 5' splice sites across introns and exons is likely important in promoting correct and efficient splicing of multi-intron pre-mRNAs.https://elifesciences.org/articles/37751splicingspliceosomesingle-moleculeRNAsnRNPCoSMoS |
| spellingShingle | Joerg E Braun Larry J Friedman Jeff Gelles Melissa J Moore Synergistic assembly of human pre-spliceosomes across introns and exons eLife splicing spliceosome single-molecule RNA snRNP CoSMoS |
| title | Synergistic assembly of human pre-spliceosomes across introns and exons |
| title_full | Synergistic assembly of human pre-spliceosomes across introns and exons |
| title_fullStr | Synergistic assembly of human pre-spliceosomes across introns and exons |
| title_full_unstemmed | Synergistic assembly of human pre-spliceosomes across introns and exons |
| title_short | Synergistic assembly of human pre-spliceosomes across introns and exons |
| title_sort | synergistic assembly of human pre spliceosomes across introns and exons |
| topic | splicing spliceosome single-molecule RNA snRNP CoSMoS |
| url | https://elifesciences.org/articles/37751 |
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