Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan.
Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.
PLoS One. 2021 Aug 18;16(8):e0256282. doi: 10.1371/journal.pone.0256282. eCollection 2021.
Human PUF-A/PUM3 is a RNA and DNA binding protein participating in the nucleolar processing of 7S to 5.8S rRNA. The nucleolar localization of PUF-A redistributes to the nucleoplasm upon the exposure to genotoxic agents in cells. However, little is known regarding the roles of PUF-A in tumor progression. Phosphoprotein database analysis revealed that Y259 phosphorylation of PUF-A is the most prevalent residue modified. Here, we reported the importance of PUF-A's phosphorylation on Y259 in tumorigenesis. PUF-A gene was knocked out by the Crispr/Cas9 method in human cervix epithelial HeLa cells. Loss of PUF-A in HeLa cells resulted in reduced clonogenic and lower transwell invasion capacity. Introduction of PUF-AY259F to PUF-A deficient HeLa cells was unable to restore colony formation. In addition, the unphosphorylated mutant of PUF-A, PUF-AY259F, attenuated PUF-A protein stability. Our results suggest the important role of Y259 phosphorylation of PUF-A in cell proliferation.
人 PUF-A/PUM3 是一种 RNA 和 DNA 结合蛋白，参与核仁加工 7S 到 5.8S rRNA。PUF-A 的核定位在细胞暴露于遗传毒性剂后重新分布到核质中。然而，关于 PUF-A 在肿瘤进展中的作用知之甚少。磷酸化蛋白数据库分析显示，PUF-A 的 Y259 磷酸化是最常见的修饰残基。在这里，我们报道了 PUF-A 的 Y259 磷酸化在肿瘤发生中的重要性。通过 Crispr/Cas9 方法在人宫颈上皮 HeLa 细胞中敲除 PUF-A 基因。在 HeLa 细胞中缺失 PUF-A 导致集落形成减少和侵袭能力降低。将 PUF-AY259F 引入 PUF-A 缺陷的 HeLa 细胞中，无法恢复集落形成。此外，PUF-A 的非磷酸化突变体 PUF-AY259F 减弱了 PUF-A 蛋白的稳定性。我们的结果表明 PUF-A 的 Y259 磷酸化在细胞增殖中起重要作用。
