Abstract
The cAMP-dependent protein kinase (PKA) pathway in Schizosaccharomyces pombe plays an important role in microtubule organization and chromosome segregation. Typically, loss of functional Pka1 induces sensitivity to the microtubule-destabilizing drug thiabendazole (TBZ) and chromosome mis-segregation. To determine the mechanism via which Pka1 is involved in these events, we explored the relevance of transcription factors by creating a double-deletion strain of pka1 and 102 individual genes encoding transcription factors. We found that rst2∆, tfs1∆, mca1∆, and moc3∆ suppressed the TBZ-sensitive phenotype of the pka1∆ strain, among which tfs1∆ was the strongest suppressor. All single mutants (rst2∆, tfs1∆, mca1∆, and moc3∆) showed a TBZ-tolerant phenotype. Tfs1 has two transcriptional domains (TFIIS and Zn finger domains), both of which contributed to the suppression of the pka1∆-induced TBZ-sensitive phenotype. pka1∆-induced chromosome mis-segregation was rescued by tfs1∆ in the presence of TBZ. tfs1 overexpression induced the TBZ-sensitive phenotype and a high frequency of chromosome mis-segregation, suggesting that the amount of Tfs1 must be strictly controlled. However, Tfs1-expression levels did not differ between the wild-type and pka1∆ strains, and the Tfs1-GFP protein was localized to the nucleus and cytoplasm in both strains, which excludes the direct regulation of expression and localization of Tfs1 by Pka1. Growth inhibition by TBZ in pka1∆ strains was notably rescued by double deletion of rst2 and tfs1 rather than single deletion of rst2 or tfs1, indicating that Rst2 and Tfs1 contribute independently to counteract TBZ toxicity. Our findings highlight Tfs1 as a key transcription factor for proper chromosome segregation.
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Acknowledgements
The authors thank all the members of our laboratory for their support and scientific advice. FY15609 (AY260-2A) was provided by the National Bio-Resource Project (NBRP), Japan. We would like to thank Editage (www.editage.com) for English language editing.
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The authors thank the faculty of Life and Environmental Sciences in Shimane University for help in financial support for publication. This work was supported by a JSPS KAKENHI Grant Number JP18K05438 (to YM) and JP19K222831 (to MK).
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KT designed the experiments, performed the experiments, made the yeast strains, and analyzed the data; SN performed the experiments, made the yeast strains, and analyzed the data; YN performed the experiments; MK analyzed the data and provided advice; YM planned the study, designed the experiments, made the plasmids and strains, performed the experiments, and analyzed the data. YM drafted the original manuscript. MK and YM reviewed and edited the original manuscript.
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Takenaka, K., Nishioka, S., Nishida, Y. et al. Tfs1, transcription elongation factor TFIIS, has an impact on chromosome segregation affected by pka1 deletion in Schizosaccharomyces pombe. Curr Genet 69, 115–125 (2023). https://doi.org/10.1007/s00294-023-01268-0
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DOI: https://doi.org/10.1007/s00294-023-01268-0