Kamel Domenick, Sookdeo Ayisha, Ikenouchi Ayana, Zhong Hualin
Department of Biological Sciences, Hunter College, The City University of New York, New York, NY, United States of America.
The Graduate Center, The City University of New York, New York, NY, United States of America.
PLoS One. 2024 Dec 12;19(12):e0312095. doi: 10.1371/journal.pone.0312095. eCollection 2024.
Nuclear pore proteins control nucleocytoplasmic transport; however, certain nucleoporins play regulatory roles in activities such as transcription and chromatin organization. The fission yeast basket nucleoporin Nup211 is implicated in mRNA export and is essential for cell viability. Nup211 preferentially associates with heterochromatin, however, it is unclear whether it plays a role in regulating transcription. To better understand its functions, we constructed a nup211 "shut-off" strain and observed that Nup211 depletion led to severe defects in cell cycle progression, including septation and cytokinesis. Using RNA-Seq and RT-qPCR, we revealed that loss of Nup211 significantly altered the mRNA levels of a set of genes crucial for cell division. Using domain analysis and CRISPR/cas9 technology, we determined that the first 655 residues of Nup211 are sufficient for viability. This truncated protein was detected at the nuclear periphery. Furthermore, exogenous expression of this domain in nup211 shut-off cells effectively restored both cell morphology and transcript abundance for some selected genes. Our findings unveil a novel role for Nup211 in regulating gene expression.
核孔蛋白控制核质运输；然而，某些核孔蛋白在转录和染色质组织等活动中发挥调节作用。裂殖酵母篮状核孔蛋白Nup211与mRNA输出有关，对细胞活力至关重要。Nup211优先与异染色质结合，然而，尚不清楚它是否在调节转录中发挥作用。为了更好地理解其功能，我们构建了一个nup211“关闭”菌株，并观察到Nup211的缺失导致细胞周期进程出现严重缺陷，包括隔膜形成和胞质分裂。使用RNA测序和逆转录定量聚合酶链反应，我们发现Nup211的缺失显著改变了一组对细胞分裂至关重要的基因的mRNA水平。通过结构域分析和CRISPR/Cas9技术，我们确定Nup211的前655个残基足以维持细胞活力。这种截短的蛋白在核周边被检测到。此外，在nup211关闭细胞中外源表达该结构域有效地恢复了一些选定基因的细胞形态和转录本丰度。我们的研究结果揭示了Nup211在调节基因表达中的新作用。
