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Large-scale remodeling of a repressed exon ribonucleoprotein to an exon definition complex active for splicing.
Elife. 2016 11 24; 5E

Abstract

Polypyrimidine-tract binding protein PTBP1 can repress splicing during the exon definition phase of spliceosome assembly, but the assembly steps leading to an exon definition complex (EDC) and how PTBP1 might modulate them are not clear. We found that PTBP1 binding in the flanking introns allowed normal U2AF and U1 snRNP binding to the target exon splice sites but blocked U2 snRNP assembly in HeLa nuclear extract. Characterizing a purified PTBP1-repressed complex, as well as an active early complex and the final EDC by SILAC-MS, we identified extensive PTBP1-modulated changes in exon RNP composition. The active early complex formed in the absence of PTBP1 proceeded to assemble an EDC with the eviction of hnRNP proteins, the late recruitment of SR proteins, and binding of the U2 snRNP. These results demonstrate that during early stages of splicing, exon RNP complexes are highly dynamic with many proteins failing to bind during PTBP1 arrest.

Authors+Show Affiliations

Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, United States.Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, United States.Department of Basic Medical Sciences, University of Arizona, Phoenix, United States.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, United States.Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, United States.Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, United States.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

27882870

Citation

Wongpalee, Somsakul Pop, et al. "Large-scale Remodeling of a Repressed Exon Ribonucleoprotein to an Exon Definition Complex Active for Splicing." ELife, vol. 5, 2016.
Wongpalee SP, Vashisht A, Sharma S, et al. Large-scale remodeling of a repressed exon ribonucleoprotein to an exon definition complex active for splicing. Elife. 2016;5.
Wongpalee, S. P., Vashisht, A., Sharma, S., Chui, D., Wohlschlegel, J. A., & Black, D. L. (2016). Large-scale remodeling of a repressed exon ribonucleoprotein to an exon definition complex active for splicing. ELife, 5. https://doi.org/10.7554/eLife.19743
Wongpalee SP, et al. Large-scale Remodeling of a Repressed Exon Ribonucleoprotein to an Exon Definition Complex Active for Splicing. Elife. 2016 11 24;5 PubMed PMID: 27882870.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Large-scale remodeling of a repressed exon ribonucleoprotein to an exon definition complex active for splicing. AU - Wongpalee,Somsakul Pop, AU - Vashisht,Ajay, AU - Sharma,Shalini, AU - Chui,Darryl, AU - Wohlschlegel,James A, AU - Black,Douglas L, Y1 - 2016/11/24/ PY - 2016/07/17/received PY - 2016/11/02/accepted PY - 2016/11/25/entrez PY - 2016/11/25/pubmed PY - 2017/11/1/medline KW - RNA binding protein KW - biochemistry KW - evolutionary biology KW - gene regulation KW - genomics KW - human KW - ribonucleoprotein KW - splicing JF - eLife JO - Elife VL - 5 N2 - Polypyrimidine-tract binding protein PTBP1 can repress splicing during the exon definition phase of spliceosome assembly, but the assembly steps leading to an exon definition complex (EDC) and how PTBP1 might modulate them are not clear. We found that PTBP1 binding in the flanking introns allowed normal U2AF and U1 snRNP binding to the target exon splice sites but blocked U2 snRNP assembly in HeLa nuclear extract. Characterizing a purified PTBP1-repressed complex, as well as an active early complex and the final EDC by SILAC-MS, we identified extensive PTBP1-modulated changes in exon RNP composition. The active early complex formed in the absence of PTBP1 proceeded to assemble an EDC with the eviction of hnRNP proteins, the late recruitment of SR proteins, and binding of the U2 snRNP. These results demonstrate that during early stages of splicing, exon RNP complexes are highly dynamic with many proteins failing to bind during PTBP1 arrest. SN - 2050-084X UR - https://www.unboundmedicine.com/medline/citation/27882870/Large_scale_remodeling_of_a_repressed_exon_ribonucleoprotein_to_an_exon_definition_complex_active_for_splicing_ L2 - https://doi.org/10.7554/eLife.19743 DB - PRIME DP - Unbound Medicine ER -