St. Vincent's Institute of Medical Research, Fitzroy, Victoria 3065, Australia.
Genetics. 2009 Aug;182(4):955-66. doi: 10.1534/genetics.109.104414. Epub 2009 Jun 1.
In yeast, assembly of the septins at the cell cortex is required for a series of key cell cycle events: bud-site selection, the morphogenesis and mitotic exit checkpoints, and cytokinesis. Here we establish that the Ccr4-Pop2-NOT mRNA deadenylase contributes to septin organization. mRNAs encoding regulators of septin assembly (Ccd42, Cdc24, Rga1, Rga2, Bem3, Gin4, Cla4, and Elm1) presented with short poly(A) tails at steady state in wild-type (wt) cells, suggesting their translation could be restricted by deadenylation. Deadenylation of septin regulators was dependent on the major cellular mRNA deadenylase Ccr4-Pop2-NOT, whereas the alternative deadenylase Pan2 played a minor role. Consistent with deadenylation of septin regulators being important for function, deletion of deadenylase subunits CCR4 or POP2, but not PAN2, resulted in septin morphology defects (e.g., ectopic bud-localized septin rings), particularly upon activation of the Cdc28-inhibitory kinase Swe1. Aberrant septin staining was also observed in the deadenylase-dead ccr4-1 mutant, demonstrating the deadenylase activity of Ccr4-Pop2 is required. Moreover, ccr4Delta, pop2Delta, and ccr4-1 mutants showed aberrant cell morphology previously observed in septin assembly mutants and exhibited genetic interactions with mutations that compromise septin assembly (shs1Delta, cla4Delta, elm1Delta, and gin4Delta). Mutations in the Not subunits of Ccr4-Pop2-NOT, which are thought to predominantly function in transcriptional control, also resulted in septin organization defects. Therefore, both mRNA deadenylase and transcriptional functions of Ccr4-Pop2-NOT contribute to septin organization in yeast.
在酵母中,细胞皮层上的 septin 组装对于一系列关键的细胞周期事件是必需的:芽位点选择、形态发生和有丝分裂退出检查点以及细胞分裂。在这里,我们确定 Ccr4-Pop2-NOT mRNA 脱腺苷酶有助于 septin 组织。在野生型 (wt) 细胞中,编码 septin 组装调节剂的 mRNAs 在稳定状态下呈现短 poly(A) 尾巴,表明它们的翻译可能受到脱腺苷化的限制。septin 调节剂的脱腺苷化依赖于主要的细胞 mRNA 脱腺苷酶 Ccr4-Pop2-NOT,而替代脱腺苷酶 Pan2 则起次要作用。与 septin 调节剂的脱腺苷化对功能很重要一致,删除脱腺苷酶亚基 CCR4 或 POP2,但不是 PAN2,会导致 septin 形态缺陷(例如,异位芽定位的 septin 环),特别是在 Cdc28 抑制激酶 Swe1 激活时。在脱腺苷酶失活的 ccr4-1 突变体中也观察到异常的 septin 染色,这表明 Ccr4-Pop2 的脱腺苷酶活性是必需的。此外,ccr4Delta、pop2Delta 和 ccr4-1 突变体表现出以前在 septin 组装突变体中观察到的异常细胞形态,并与破坏 septin 组装的突变体表现出遗传相互作用(shs1Delta、cla4Delta、elm1Delta 和 gin4Delta)。Ccr4-Pop2-NOT 的 Not 亚基中的突变,这些突变被认为主要在转录控制中起作用,也导致 septin 组织缺陷。因此,Ccr4-Pop2-NOT 的 mRNA 脱腺苷酶和转录功能都有助于酵母中 septin 的组织。