Cwc23, an essential J protein critical for pre-mRNA splicing with a dispensable J domain

Mol Cell Biol. 2010 Jan;30(1):33-42. doi: 10.1128/MCB.00842-09.

Abstract

J proteins are structurally diverse, obligatory cochaperones of Hsp70s, each with a highly conserved J domain that plays a critical role in the stimulation of Hsp70's ATPase activity. The essential protein, Cwc23, is one of 13 J proteins found in the cytosol and/or nucleus of Saccharomyces cerevisiae. We report that a partial loss-of-function CWC23 mutant has severe, global defects in pre-mRNA splicing. This mutation leads to accumulation of the excised, lariat form of the intron, as well as unspliced pre-mRNA, suggesting a role for Cwc23 in spliceosome disassembly. Such a role is further supported by the observation that this mutation results in reduced interaction between Cwc23 and Ntr1 (SPP382), a known component of the disassembly pathway. However, Cwc23 is a very atypical J protein. Its J domain, although functional, is dispensable for both cell viability and pre-mRNA splicing. Nevertheless, strong genetic interactions were uncovered between point mutations encoding alterations in Cwc23's J domain and either Ntr1 or Prp43, a DExD/H-box helicase essential for spliceosome disassembly. These genetic interactions suggest that Hsp70-based chaperone machinery does play a role in the disassembly process. Cwc23 provides a unique example of a J protein; its partnership with Hsp70 plays an auxiliary, rather than a central, role in its essential cellular function.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Survival / physiology
  • DEAD-box RNA Helicases / physiology
  • MicroRNAs / physiology*
  • Molecular Chaperones / genetics
  • Molecular Chaperones / physiology*
  • Point Mutation
  • Protein Structure, Tertiary
  • RNA Splicing*
  • RNA, Fungal / physiology*
  • RNA, Messenger / physiology*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / physiology*
  • Spliceosomes / physiology

Substances

  • Cwc23 protein, S cerevisiae
  • MicroRNAs
  • Molecular Chaperones
  • RNA, Fungal
  • RNA, Messenger
  • Saccharomyces cerevisiae Proteins
  • Spp382 protein, S cerevisiae
  • PRP43 protein, S cerevisiae
  • DEAD-box RNA Helicases