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Functional interactions between Prp8, Prp18, Slu7, and U5 snRNA during the second step of pre-mRNA splicing

Anna Aronova1, Dagmar Bacíková, Luciana B Crotti

  • 1Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10021, USA.

RNA (New York, N.Y.)
|July 14, 2007
PubMed

Insights

Mutations in the spliceosome protein Prp8 can cause temperature-sensitive growth defects. Suppressing these mutations reveals a functional link between Prp8 and splicing factors Prp18 and Slu7.

Area of Science:

  • Molecular Biology
  • RNA Splicing Mechanisms

Background:

  • The Prp22 helicase releases spliced mRNA after the second transesterification step.
  • Prp8 protein interactions within the spliceosome are critical for this process.
  • Mutations in Prp8 at Arg1753 suppress defects in Prp22 and cause temperature sensitivity.

Purpose of the Study:

  • Investigate the role of Prp8-R1753 interactions in pre-mRNA splicing.
  • Identify factors that suppress temperature-sensitive mutations in Prp8.
  • Elucidate the functional relationship between Prp8 and other splicing factors.

Main Methods:

  • Genetic suppression analysis using temperature-sensitive mutants.
  • In vitro pre-mRNA splicing assays.
  • Analysis of U5 snRNA and Prp18 mutations.

Main Results:

  • Mutations in U5 snRNA loop I or in Prp18 suppress temperature-sensitive prp8-R1753 mutants.
  • Gain-of-function Prp18 alleles also alleviate growth defects in slu7 mutants.
  • Prp8-R1753 mutations impair step 2 of pre-mRNA splicing in vitro.

Conclusions:

  • Arg1753 in Prp8 stabilizes U5/exon interactions before exon joining.
  • These interactions are maintained until disrupted by the Prp22 helicase.
  • A functional link exists between Prp8, Prp18, and Slu7 in the second step of splicing.

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