Department of Parasitology, Institute of Biomedical Sciences, Sāo Paulo University (USP), Sao Paulo, Brazil.
Department of Bioprocess and Biotechnology, School of Agriculture, Sao Paulo State University (UNESP), Botucatu, Brazil.
PLoS Comput Biol. 2022 May 19;18(5):e1010081. doi: 10.1371/journal.pcbi.1010081. eCollection 2022 May.
Ethanol alters many subsystems of Saccharomyces cerevisiae, including the cell cycle. Two ethanol-responsive lncRNAs in yeast interact with cell cycle proteins, and here, we investigated the role of these RNAs in cell cycle. Our network dynamic modeling showed that higher and lower ethanol-tolerant strains undergo cell cycle arrest in mitosis and G1 phases, respectively, during ethanol stress. The higher population rebound of the lower ethanol-tolerant phenotype after stress relief responds to the late phase arrest. We found that the lncRNA lnc9136 of SEY6210 (a lower ethanol-tolerant strain) induces cells to skip mitosis arrest. Simulating an overexpression of lnc9136 and analyzing CRISPR-Cas9 mutants lacking this lncRNA suggest that lnc9136 induces a regular cell cycle even under ethanol stress, indirectly regulating Swe1p and Clb1/2 by binding to Gin4p and Hsl1p. Notably, lnc10883 of BY4742 (a higher ethanol-tolerant strain) does not prevent G1 arrest in this strain under ethanol stress. However, lnc19883 circumvents DNA and spindle damage checkpoints, maintaining a functional cell cycle by interacting with Mec1p or Bub1p even in the presence of DNA/spindle damage. Overall, we present the first evidence of direct roles for lncRNAs in regulating yeast cell cycle proteins, the dynamics of this system in different ethanol-tolerant phenotypes, and a new yeast cell cycle model.
乙醇改变了酿酒酵母的许多子系统，包括细胞周期。酵母中两种对乙醇有反应的 lncRNA 与细胞周期蛋白相互作用，在这里，我们研究了这些 RNA 在细胞周期中的作用。我们的网络动态模型表明，在乙醇胁迫下，较高和较低乙醇耐受菌株分别在有丝分裂和 G1 期停滞细胞周期。应激缓解后较低乙醇耐受表型的较高种群反弹对应于后期停滞。我们发现 SEY6210（较低乙醇耐受菌株）的 lncRNA lnc9136 使细胞跳过有丝分裂停滞。模拟 lnc9136 的过表达并分析缺乏这种 lncRNA 的 CRISPR-Cas9 突变体表明，lnc9136 甚至在乙醇胁迫下诱导正常的细胞周期，通过与 Gin4p 和 Hsl1p 结合间接调节 Swe1p 和 Clb1/2。值得注意的是，BY4742（较高乙醇耐受菌株）的 lnc10883 不能阻止该菌株在乙醇胁迫下的 G1 期停滞。然而，lnc19883 绕过 DNA 和纺锤体损伤检查点，通过与 Mec1p 或 Bub1p 相互作用，即使在存在 DNA/纺锤体损伤的情况下，也能维持功能正常的细胞周期。总的来说，我们首次提供了 lncRNA 直接调节酵母细胞周期蛋白的证据，不同乙醇耐受表型中该系统的动力学，以及一个新的酵母细胞周期模型。
