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<p>Alternative splicing of precursor-messenger RNAs (pre-mRNA) bearing introns with competing 5’ splice sites (5’SS) produces two mRNAs. We investigated the mechanism of alternative splicing of the yeast <i>HEH1/SRC1</i> gene, which contains four nucleotide-apart competing 5’SS, by monitoring its RNA and protein products. <i>HEH1</i> alternative splicing requires a sixteen-nucleotide pre-mRNA segment spanning its two 5’SS. The nucleotides are decoded by U5 and U6 small nuclear RNAs (snRNAs), supported by specific proteins of the spliceosomal B and Bact complexes, including Prp8. <i>HEH1</i> alternative splicing became independent of the supporting proteins following recalibration of the pre-mRNA-snRNA base pairings through changes in the 5’SS or U5 and U6 snRNAs. Assisted by proteins that stabilize low-fidelity, high-efficiency spliceosome conformations, the competing <i>HEH1</i> 5’SS are marked for alternative splicing by U5 and U6 snRNAs during the B to Bact transition of the spliceosome.</p>

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Mechanism of alternative splicing of yeast HEH1 through competing 5’ splice sites

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月16日

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  • Ankita Katoch Banyal,
  • Poulami Choudhuri,
  • Balashankar R. Pillai,
  • Amjadudheen Varikkapulakkal,
  • Shravan Kumar Mishra

摘要

Alternative splicing of precursor-messenger RNAs (pre-mRNA) bearing introns with competing 5’ splice sites (5’SS) produces two mRNAs. We investigated the mechanism of alternative splicing of the yeast HEH1/SRC1 gene, which contains four nucleotide-apart competing 5’SS, by monitoring its RNA and protein products. HEH1 alternative splicing requires a sixteen-nucleotide pre-mRNA segment spanning its two 5’SS. The nucleotides are decoded by U5 and U6 small nuclear RNAs (snRNAs), supported by specific proteins of the spliceosomal B and Bact complexes, including Prp8. HEH1 alternative splicing became independent of the supporting proteins following recalibration of the pre-mRNA-snRNA base pairings through changes in the 5’SS or U5 and U6 snRNAs. Assisted by proteins that stabilize low-fidelity, high-efficiency spliceosome conformations, the competing HEH1 5’SS are marked for alternative splicing by U5 and U6 snRNAs during the B to Bact transition of the spliceosome.