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Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
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A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
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The tumour suppressor RBM5 activates the helicase DHX15 to regulate splicing.

Z Hong Zhou1,2, Shiheng Liu1, Tiantian Su1

  • 1Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095, USA.

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Summary

The tumor suppressor RBM5 acts as a spliceosome gatekeeper, controlling apoptosis-regulating splicing. It physically blocks spliceosome progression while activating the DHX15 helicase for tumor-suppressive alternative splicing.

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Area of Science:

  • Molecular Biology
  • Structural Biology
  • Cancer Research

Background:

  • Pre-mRNA splicing dictates the proteome and is often altered in cancer.
  • The tumor suppressor RBM5 regulates an exon network involved in apoptosis, but its mechanism is unclear.

Purpose of the Study:

  • To elucidate the molecular mechanism of RBM5 in regulating pre-mRNA splicing.
  • To determine the structural basis of RBM5's interaction with the spliceosome.
  • To understand how RBM5's function is linked to cancer-associated mutations.

Main Methods:

  • In vivo spliceosome capture coupled with cryogenic electron microscopy (cryo-EM).
  • Structural determination of spliceosomes arrested by RBM5.
  • Functional assays to test the impact of disrupting RBM5 interaction interfaces.

Main Results:

  • RBM5 binds the SF3B1 HEAT surface of the spliceosome.
  • RBM5 acts as a dual gatekeeper: sterically inhibiting spliceosome progression and activating the DHX15 helicase.
  • RBM5 interfaces with DHX15 and SF3B1 are crucial for its exon repression function; disruption inhibits this activity.

Conclusions:

  • RBM5 functions as a dual-action spliceosome gatekeeper, coupling helicase activation with physical stalling.
  • This mechanism enforces tumor-suppressive alternative splicing programs.
  • Cancer-associated mutations at RBM5 regulatory interfaces may disrupt its tumor-suppressive role in apoptosis regulation.