Characterization of Cwc2, U6 snRNA, and Prp8 interactions destabilized by Prp16 ATPase at the transition between the first and second steps of splicing.

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  • Additional Information
    • Source:
      Publisher: Cold Spring Harbor Laboratory Press Country of Publication: United States NLM ID: 9509184 Publication Model: Electronic Cited Medium: Internet ISSN: 1469-9001 (Electronic) Linking ISSN: 13558382 NLM ISO Abbreviation: RNA Subsets: MEDLINE
    • Publication Information:
      Publication: <2003->: Cold Spring Harbor, NY : Cold Spring Harbor Laboratory Press
      Original Publication: New York, NY : Cambridge University Press, c1995-
    • Subject Terms:
      Saccharomyces cerevisiae Proteins*/genetics ; Saccharomyces cerevisiae Proteins*/metabolism ; Saccharomyces cerevisiae Proteins*/chemistry ; RNA, Small Nuclear*/genetics ; RNA, Small Nuclear*/metabolism ; RNA, Small Nuclear*/chemistry ; RNA Splicing* ; Saccharomyces cerevisiae*/genetics ; Saccharomyces cerevisiae*/metabolism ; Spliceosomes*/metabolism ; Spliceosomes*/genetics ; RNA Splicing Factors*/metabolism ; RNA Splicing Factors*/genetics ; RNA Splicing Factors*/chemistry; Introns/genetics ; Ribonucleoprotein, U4-U6 Small Nuclear/metabolism ; Ribonucleoprotein, U4-U6 Small Nuclear/genetics ; Ribonucleoprotein, U4-U6 Small Nuclear/chemistry ; Cryoelectron Microscopy ; Mutation ; Protein Binding ; Catalytic Domain ; Alleles ; DEAD-box RNA Helicases/metabolism ; DEAD-box RNA Helicases/genetics ; DEAD-box RNA Helicases/chemistry ; Adenosine Triphosphatases/metabolism ; Adenosine Triphosphatases/genetics ; Adenosine Triphosphatases/chemistry ; RNA-Binding Proteins ; Ribonucleoprotein, U5 Small Nuclear ; RNA Helicases
    • Abstract:
      The spliceosome performs two consecutive transesterification reactions using one catalytic center, thus requiring its rearrangement between the two catalytic steps of splicing. The Prp16 ATPase facilitates exit from the first-step conformation of the catalytic center by destabilizing some interactions important for catalysis. To better understand rearrangements within the Saccharomyces cerevisiae catalytic center, we characterize factors that modulate the function of Prp16: Cwc2, N-terminal domain of Prp8, and U6- 41 AACAAU 46 region. Alleles of these factors were identified through genetic screens for mutants that correct cs defects of prp16-302 alleles. Several of the identified U6, cwc2 , and prp8 alleles are located in close proximity of each other in cryo-EM structures of the spliceosomal catalytic conformations. Cwc2 and U6 interact with the intron sequences in the first step, but they do not seem to contribute to the stability of the second-step catalytic center. On the other hand, the N-terminal segment of Prp8 not only affects intron positioning for the first step, but it also makes important contacts in the proximity of the active site for both the first and second steps of splicing. By identifying interactions important for the stability of catalytic conformations, our genetic analyses indirectly inform us about features of the transition-state conformation of the spliceosome.
      (© 2024 Meissner et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)
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    • Contributed Indexing:
      Keywords: Prp16; Prp8; U6 snRNA; spliceosomal catalytic center; transition-state conformation
    • Accession Number:
      0 (Saccharomyces cerevisiae Proteins)
      0 (RNA, Small Nuclear)
      0 (PRP8 protein, S cerevisiae)
      0 (U6 small nuclear RNA)
      EC 3.6.1.- (PRP16 protein, S cerevisiae)
      0 (RNA Splicing Factors)
      0 (Cwc2 protein, S cerevisiae)
      0 (Ribonucleoprotein, U4-U6 Small Nuclear)
      EC 3.6.4.13 (DEAD-box RNA Helicases)
      EC 3.6.1.- (Adenosine Triphosphatases)
      0 (RNA-Binding Proteins)
      0 (Ribonucleoprotein, U5 Small Nuclear)
      EC 3.6.4.13 (RNA Helicases)
    • Publication Date:
      Date Created: 20240614 Date Completed: 20240816 Latest Revision: 20240902
    • Publication Date:
      20240902
    • Accession Number:
      PMC11331412
    • Accession Number:
      10.1261/rna.079886.123
    • Accession Number:
      38876504