Fujii Shiori, Duy Duong Long, Valderrama Arvin Lapiz, Takeuchi Risa, Matsuura Eri, Ito Ayaka, Irie Kaoru, Suda Yasuyuki, Mizuno Tomoaki, Irie Kenji
Department of Molecular Cell Biology, Graduate School of Comprehensive Human Sciences and Faculty of Medicine, University of Tsukuba, Tsukuba, Japan.
Department of Molecular Cell Biology, Graduate School of Comprehensive Human Sciences and Faculty of Medicine, University of Tsukuba, Tsukuba, Japan; Ph.D. Program in Human Biology, School of Integrative and Global Majors, University of Tsukuba, Tsukuba, Japan.
Biochem Biophys Res Commun. 2021 Sep 17;570:125-130. doi: 10.1016/j.bbrc.2021.07.007. Epub 2021 Jul 16.
There are two major deadenylase complexes, Ccr4-Not and Pan2-Pan3, which shorten the 3' poly(A) tail of mRNA and are conserved from yeast to human. We have previously shown that the Ccr4-mediated deadenylation plays the important role in gene expression regulation in the yeast stationary phase cell. In order to further understand the role of deadenylases in different growth condition, in this study we investigated the effect of deletion of both deadenylases on the cell in non-fermentable carbon containing media. We found that both ccr4Δ and ccr4Δ pan2Δ mutants showed similar growth defect in YPD media: when switched to media containing non-fermentable source (Glycerol-Lactate) only the ccr4Δ grew while the ccr4Δ pan2Δ did not. Ccr4, Pan2, and Pan3 were phosphorylated in GlyLac medium, suggesting that the activities of Ccr4, Pan2, and Pan3 may be regulated by phosphorylation in response to change of carbon sources. To get insights how Ccr4 and Pan2 function in the cell growth in media containing non-fermentable source only, we isolated multicopy suppressors for the growth defect on YPGlyLac media of the ccr4Δ pan2Δ mutant and identified two genes, STM1 and REX2, which encode a ribosome-associated protein and a 3'-5' RNA exonuclease, respectively. Our results suggest that the Pan2-Pan3 complex, together with the Ccr4-Not complex, has important roles in the growth on non-fermentable carbon sources.
有两种主要的去腺苷酸化酶复合物，即Ccr4-Not和Pan2-Pan3，它们会缩短mRNA的3'多聚腺苷酸尾巴，并且从酵母到人类都是保守的。我们之前已经表明，Ccr4介导的去腺苷酸化在酵母静止期细胞的基因表达调控中起重要作用。为了进一步了解去腺苷酸化酶在不同生长条件下的作用，在本研究中，我们研究了缺失这两种去腺苷酸化酶对细胞在含非发酵碳源培养基中的影响。我们发现，ccr4Δ和ccr4Δ pan2Δ突变体在YPD培养基中表现出相似的生长缺陷：当切换到含有非发酵源（甘油-乳酸）的培养基时，只有ccr4Δ能够生长，而ccr4Δ pan2Δ不能。Ccr4、Pan2和Pan3在GlyLac培养基中被磷酸化，这表明Ccr4、Pan2和Pan3的活性可能通过磷酸化来响应碳源的变化而受到调节。为了深入了解Ccr4和Pan2在仅含非发酵源的培养基中细胞生长中的作用，我们分离了ccr4Δ pan2Δ突变体在YPGlyLac培养基上生长缺陷的多拷贝抑制子，并鉴定了两个基因STM1和REX2，它们分别编码一种核糖体相关蛋白和一种3'-5'RNA外切核酸酶。我们的结果表明，Pan2-Pan3复合物与Ccr4-Not复合物一起，在利用非发酵碳源生长中起重要作用。
