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    Synergistic roles for human U1 snRNA stem-loops in pre-mRNA splicing

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    Name:
    Martelly_Sharma_Manuscript+Fig ...
    Embargo:
    2022-06-09
    Size:
    30.28Mb
    Format:
    PDF
    Description:
    Final Accepted Manuscript
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    SupplementaryDataFile_Tables.zip
    Embargo:
    2022-06-09
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    Author
    Martelly, William
    Fellows, Bernice
    Kang, Paul
    Vashisht, Ajay
    Wohlschlegel, James A.
    Sharma, Shalini
    Affiliation
    Department of Basic Medical Sciences, College of Medicine-Phoenix, University of Arizona
    Department of Epidemiology and Biostatistics, Mel and Enid Zuckerman College of Public Health-Phoenix, University of Arizona
    Issue Date
    2021-06-09
    Keywords
    A complex
    DDX39A
    DDX39B
    E complex
    SF3A1
    Spliceosome
    Stem-loop
    U1 snRNA
    U1 snRNP
    U2 snRNP
    UAP56
    URH49
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    Metadata
    Show full item record
    Publisher
    Informa UK Limited
    Citation
    Martelly, W., Fellows, B., Kang, P., Vashisht, A., Wohlschlegel, J. A., & Sharma, S. (2021). Synergistic roles for human U1 snRNA stem-loops in pre-mRNA splicing. RNA Biology.
    Journal
    RNA Biology
    Rights
    © 2021 Informa UK Limited, trading as Taylor & Francis Group.
    Collection Information
    This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at repository@u.library.arizona.edu.
    Abstract
    During spliceosome assembly, interactions that bring the 5′ and 3′ ends of an intron in proximity are critical for the production of mature mRNA. Here, we report synergistic roles for the stem-loops 3 (SL3) and 4 (SL4) of the human U1 small nuclear RNA (snRNA) in maintaining the optimal U1 snRNP function, and formation of cross-intron contact with the U2 snRNP. We find that SL3 and SL4 bind distinct spliceosomal proteins and combining a U1 snRNA activity assay with siRNA-mediated knockdown, we demonstrate that SL3 and SL4 act through the RNA helicase UAP56 and the U2 protein SF3A1, respectively. In vitro analysis using UV crosslinking and splicing assays indicated that SL3 likely promotes the SL4-SF3A1 interaction leading to enhancement of A complex formation and pre-mRNA splicing. Overall, these results highlight the vital role of the distinct contacts of SL3 and SL4 in bridging the pre-mRNA bound U1 and U2 snRNPs during the early steps of human spliceosome assembly. © 2021 Informa UK Limited, trading as Taylor & Francis Group.
    Note
    12 month embargo; published online: 09 June 2021
    ISSN
    1547-6286
    EISSN
    1555-8584
    DOI
    10.1080/15476286.2021.1932360
    Version
    Final accepted manuscript
    Sponsors
    National Institute of General Medical Sciences
    ae974a485f413a2113503eed53cd6c53
    10.1080/15476286.2021.1932360
    Scopus Count
    Collections
    UA Faculty Publications

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