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2013 Fiscal Year Final Research Report

Analysis of the assembly and functional defect of the spliceosome associated with Retinitis pigmentosa

Research Project

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Project/Area Number 23790068
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Biological pharmacy
Research InstitutionHokkaido University

Principal Investigator

MAITA Hiroshi  北海道大学, 薬学研究科(研究院), 講師 (60431318)

Project Period (FY) 2011 – 2012
Keywords分子生物学 / スプライシング / snRNP
Research Abstract

The spliceosome is a highly dynamic macromolecular ribonucleoprotein (RNP) machine that catalyzes pre-mRNA splicing by assembling U1, U2, U4, U5, and U6 small nuclear (sn)RNPs. To process large numbers of introns, synthesis and recycling of snRNPs must be maintained within an appropriate range to avoid their shortage, which would cause a severe disease such as Retinitis pigmentosa. However, the mechanism that maintains cellular snRNP levels is unknown. We constructed an expression library of a luciferase fragment fused to core components of snRNPs and used it to isolate PRPF6 and U5-40K that specifically reconstitute luciferase activity in the U5 snRNP complex. We revealed that the reporter detects the effects of small molecules on the levels of the U5 snRNP-reporter protein complex. Our assay will be useful to identify the small molecule that can control the snRNP level in an appropriate range.

  • Research Products

    (4 results)

All 2014 2012 2011

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Presentation (3 results)

  • [Journal Article] A split luciferase-based reporter for detection of a cellular macromolecular complex2014

    • Author(s)
      Maita H., Tomita K. and Ariga H
    • Journal Title

      Anal. Biochem

      Volume: 452 Pages: 1-9

    • Peer Reviewed
  • [Presentation] Detection of Cellular U5 snRNP Levels Using Split Luciferase2014

    • Author(s)
      Maita H. and Ariga H
    • Organizer
      The 16^<th> Annual Meeting of the RNA Society
    • Place of Presentation
      Kyoto, Japan
    • Year and Date
      20140614-18
  • [Presentation] Analysis of the spliceosome formation by a newly developed snRNP reporter2012

    • Author(s)
      Tomita K., Ariga H. and Maita H
    • Organizer
      日本分子生物学会年会
    • Place of Presentation
      福岡
    • Year and Date
      2012-12-13
  • [Presentation] A possible role for Prp3-Sad1/USP39 interaction in the spliceosomal assembly2011

    • Author(s)
      Shiida M., Ariga H. and Maita H
    • Organizer
      日本分子生物学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2011-12-14

URL: 

Published: 2015-06-25  

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