W.M. Keck Science Center, The Claremont Colleges, Claremont, CA 91711, USA.
Exp Cell Res. 2012 Oct 1;318(16):2071-84. doi: 10.1016/j.yexcr.2012.05.020. Epub 2012 Jun 6.
Schizosaccharomyces pombe Dsk1 is an SR protein-specific kinase (SRPK), whose homologs have been identified in every eukaryotic organism examined. Although discovered as a mitotic regulator with protein kinase activity toward SR splicing factors, it remains largely unknown about what and how Dsk1 contributes to cell cycle and pre-mRNA splicing. In this study, we investigated the Dsk1 function by determining interacting factors and cellular localization of the kinase. Consistent with its reported functions, we found that pre-mRNA processing and cell cycle factors are prominent among the proteins co-purified with Dsk1. The identification of these factors led us to find Rsd1 as a novel Dsk1 substrate, as well as the involvement of Dsk1 in cellular distribution of poly(A)(+) RNA. In agreement with its role in nuclear events, we also found that Dsk1 is mainly localized in the nucleus during G(2) phase and at mitosis. Furthermore, we revealed the oscillation of Dsk1 protein in a cell cycle-dependent manner. This paper marks the first comprehensive analysis of in vivo Dsk1-associated proteins in fission yeast. Our results reflect the conserved role of SRPK family in eukaryotic organisms, and provide information about how Dsk1 functions in pre-mRNA processing and cell-division cycle.
裂殖酵母 Dsk1 是一种富含丝氨酸/精氨酸的剪接因子特异性蛋白激酶(SRPK),其同源物已在所有被研究的真核生物中被鉴定出来。尽管 Dsk1 最初被发现作为一种有丝分裂调节因子,具有针对 SR 剪接因子的蛋白激酶活性,但关于 Dsk1 如何参与细胞周期和前体 mRNA 剪接的功能仍然知之甚少。在这项研究中,我们通过确定激酶的相互作用因子和细胞定位来研究 Dsk1 的功能。与已报道的功能一致,我们发现前体 mRNA 处理和细胞周期因子是与 Dsk1 共纯化的蛋白质中的主要成分。这些因子的鉴定使我们发现了 Rsd1 是 Dsk1 的一种新的底物,以及 Dsk1 参与多聚(A)+ RNA 的细胞分布。与它在核事件中的作用一致,我们还发现 Dsk1 在 G2 期和有丝分裂期间主要定位于细胞核中。此外,我们还揭示了 Dsk1 蛋白在细胞周期依赖性的方式下呈现出振荡。本文标志着首次对裂殖酵母中 Dsk1 相关蛋白进行全面的体内分析。我们的结果反映了 SRPK 家族在真核生物中的保守作用,并提供了关于 Dsk1 在前体 mRNA 处理和细胞分裂周期中发挥作用的信息。