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Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
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SRSF3 Is a Critical Requirement for Inclusion of Exon 3 of BIS Pre-mRNA

Ji-Ye Baek1,2,3, Hye-Hyeon Yun1,2, Soon-Young Jung1,2

  • 1Department of Biochemistry, The Catholic University of Korea, 222, Banpo-daero, Seocho-gu, Seoul 06591, Korea.

Cells
|October 22, 2020
PubMed

Insights

The study identifies novel messenger RNA (mRNA) isomers of BCL-2 interacting cell death suppressor (BIS), also known as BAG3, revealing that SRSF3 protein regulates BIS splicing. This regulation is crucial for maintaining BIS protein integrity and nuclear translocation of HSF1 during cellular stress.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Cellular Stress Response

Background:

  • BCL-2 interacting cell death suppressor (BIS), or BAG3, is a key protein implicated in cancer and myopathy.
  • Limited research exists on the alternative splicing of BIS pre-messenger RNA (pre-mRNA).

Purpose of the Study:

  • To investigate the alternative splicing of BIS pre-mRNA and identify novel mRNA isomers.
  • To elucidate the role of SRSF3 in regulating BIS pre-mRNA splicing and its functional consequences.

Main Methods:

  • RT-PCR and sequencing were employed to identify BIS mRNA isomers in various cell lines and mouse tissues.
  • SRSF3 depletion experiments were conducted using endogenous BIS and a GFP-BIS minigene.
  • Analysis of HSF1 nuclear translocation was performed in response to SRSF3 depletion and BIS variants.

Main Results:

  • Novel BIS mRNA isomers, with exon 3 or exons 2-3 skipped, were identified.
  • SRSF3 depletion significantly promoted exon 3 skipping of BIS pre-mRNA.
  • SRSF3 binds to exon 3, and its deletion mimics the skipping effect of SRSF3 knockdown.
  • SRSF3 depletion led to a truncated BIS protein, inhibiting HSF1 nuclear translocation, which was rescued by wild-type BIS but not exon 3-deleted BIS.

Conclusions:

  • SRSF3 plays a critical role in maintaining the correct splicing of BIS pre-mRNA, preventing exon 3 skipping.
  • The integrity of the BIS protein, regulated by SRSF3-mediated splicing, is essential for proper HSF1 localization under cellular stress conditions.
Keywords:
BISHSF1SRSF3splicing

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