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全局基因表达分析裂殖酵母突变体中 RNA pol II C 端结构域丝氨酸 2 位磷酸化缺陷。

Global gene expression analysis of fission yeast mutants impaired in Ser-2 phosphorylation of the RNA pol II carboxy terminal domain.

机构信息

Department of Biology, University of Western Ontario, London, Ontario, Canada.

出版信息

PLoS One. 2011;6(9):e24694. doi: 10.1371/journal.pone.0024694. Epub 2011 Sep 12.

Abstract

In Schizosaccharomyces pombe the nuclear-localized Lsk1p-Lsc1p cyclin dependent kinase complex promotes Ser-2 phosphorylation of the heptad repeats found within the RNA pol II carboxy terminal domain (CTD). Here, we first provide evidence supporting the existence of a third previously uncharacterized Ser-2 CTD kinase subunit, Lsg1p. As expected for a component of the complex, Lsg1p localizes to the nucleus, promotes Ser-2 phosphorylation of the CTD, and physically interacts with both Lsk1p and Lsc1p in vivo. Interestingly, we also demonstrate that lsg1Δ mutants--just like lsk1Δ and lsc1Δ strains--are compromised in their ability to faithfully and reliably complete cytokinesis. Next, to address whether kinase mediated alterations in CTD phosphorylation might selectively alter the expression of genes with roles in cytokinesis and/or the cytoskeleton, global gene expression profiles were analyzed. Mutants impaired in Ser-2 phosphorylation display little change with respect to the level of transcription of most genes. However, genes affecting cytokinesis--including the actin interacting protein gene, aip1--as well as genes with roles in meiosis, are included in a small subset that are differentially regulated. Significantly, genetic analysis of lsk1Δ aip1Δ double mutants is consistent with Lsk1p and Aip1p acting in a linear pathway with respect to the regulation of cytokinesis.

摘要

在裂殖酵母中,定位于核内的 Lsk1p-Lsc1p 细胞周期蛋白依赖性激酶复合物促进 RNA pol II CTD 内七肽重复序列的 Ser-2 磷酸化。在这里,我们首先提供了支持存在第三个以前未表征的 Ser-2 CTD 激酶亚基 Lsg1p 的证据。与复合物的组成部分一样,Lsg1p 定位于细胞核内,促进 CTD 的 Ser-2 磷酸化,并在体内与 Lsk1p 和 Lsc1p 相互作用。有趣的是,我们还证明 lsg1Δ 突变体——就像 lsk1Δ 和 lsc1Δ 菌株一样——在准确可靠地完成胞质分裂的能力上受到了损害。接下来,为了解激酶介导的 CTD 磷酸化改变是否可能选择性地改变在胞质分裂和/或细胞骨架中起作用的基因的表达,我们分析了全基因组基因表达谱。在 Ser-2 磷酸化中受损的突变体,与大多数基因的转录水平相比,变化很小。然而,影响胞质分裂的基因——包括肌动蛋白相互作用蛋白基因 aip1——以及在减数分裂中起作用的基因,都被包含在一个小的差异调控亚群中。重要的是,lsk1Δ aip1Δ 双突变体的遗传分析表明,Lsk1p 和 Aip1p 在调节胞质分裂方面呈线性途径作用。

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