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A conformational rearrangement in the spliceosome sets the stage for Prp22-dependent mRNA release

Beate Schwer1

  • 1Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY 10065, USA. bschwer@med.cornell.edu

Molecular Cell
|June 24, 2008
PubMed

Insights

The DEAH box RNA helicase Prp22 binds mRNA downstream of the exon-exon junction. This interaction is crucial for disrupting spliceosome contacts and releasing the mRNA product after splicing.

Area of Science:

  • Molecular Biology
  • RNA processing
  • Gene expression

Background:

  • Pre-mRNA splicing is a fundamental process for gene expression, producing mature mRNA from precursor transcripts.
  • The spliceosome, a large molecular machine, catalyzes splicing, but its disassembly and product release are complex.
  • The DEAH box RNA helicase Prp22 is known to be involved in mRNA release from the spliceosome.

Purpose of the Study:

  • To elucidate the mechanism by which Prp22 facilitates mRNA release from the spliceosome.
  • To identify the specific interaction site of Prp22 on the mRNA.
  • To understand the role of Prp22 in remodeling spliceosome-U5 snRNP contacts.

Main Methods:

  • Site-specific crosslinking experiments to map RNA-protein interactions.
  • RNase H protection assays to assess RNA accessibility and binding.
  • Biochemical assays to study spliceosome disassembly and mRNA release.

Main Results:

  • Prp22 directly interacts with the mRNA downstream of the exon-exon junction prior to mRNA release.
  • This interaction occurs after the second transesterification step of splicing.
  • Prp22 binding leads to the disruption of contacts between the mRNA and the U5 small nuclear ribonucleoprotein (snRNP).

Conclusions:

  • Prp22 acts as a 3' to 5' helicase that targets the mRNA downstream of the exon-exon junction.
  • By remodeling mRNA/U5 snRNP interactions, Prp22 facilitates the liberation of the spliced mRNA from the spliceosome.
  • These findings provide a mechanistic insight into the essential step of mRNA release during pre-mRNA splicing.

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