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bioRxiv · 10.1101/2025.09.02.673742

Differential quantitative requirements for pre-mRNA splicing-regulated shelterin protein levels in distinct telomere functions

Takeuchi, M.·Otsubo, Y.·Kanoh, J.
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Abstract

Telomeres perform multiple functions to maintain genome stability, including telomere length regulation, chromosome end protection, and meiotic chromosome dynamics. These functions are governed by shelterin, a telomere-binding protein complex. Here, we show that efficient pre-mRNA splicing of the Schizosaccharomyces pombe shelterin components Rap1 and Poz1 ensures sufficient protein levels, which are critical for telomere maintenance. Our analyses revealed that Tls1 and Cay1 act at distinct steps in splicing, specifically affecting rap1 and poz1 transcripts: Tls1 strongly interacts with Brr2 (a splicing factor), whereas Cay1 preferentially associates with introns. Accordingly, deletion of tls1 and cay1 synergistically impaired splicing of rap1 and poz1 transcripts and reduced their protein levels, leading to abnormal telomere elongation. Removal of introns from the rap1 and poz1 genes restored normal protein levels and telomere length, confirming that defective splicing underlies these defects. Analyses of the phenotypes of single and double tls1{Delta} and cay1{Delta} mutants revealed that different telomere functions vary in their dependence on Rap1 levels: telomere length regulation and, to a lesser extent, meiosis require higher protein abundance, whereas chromosome end protection can be sustained with minimal amounts. These findings reveal a hierarchical requirement for Rap1 across telomere functions and establish a framework for understanding how splicing-dependent regulation of shelterin components coordinates multiple aspects of telomere biology.

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Takeuchi, M., Otsubo, Y., Kanoh, J.. 2025-09-02. Differential quantitative requirements for pre-mRNA splicing-regulated shelterin protein levels in distinct telomere functions. https://doi.org/10.1101/2025.09.02.673742

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