The RNA binding domain of Pumilio antagonizes poly-adenosine binding protein and accelerates deadenylation.
The RNA binding domain of Pumilio antagonizes poly-adenosine binding protein and accelerates deadenylation.
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DOI：
10.1261/rna.046029.114
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发表时间：
2014-08
期刊：
影响因子：
--
通讯作者：
Goldstrohm AC
中科院分区：
文献类型：
--
作者：
Weidmann CA;Raynard NA;Blewett NH;Van Etten J;Goldstrohm AC
本文分析了Pumilio抑制靶基因翻译的机制。结果显示，相当令人惊讶的是，去腺苷化的促进不是表达所必需的。相反，Pumilio与poly（A）结合蛋白相互作用，并以某种方式干扰其活性。PUF蛋白是在干细胞维持、神经过程和胚胎发育中起重要作用的有效阻遏物。这些功能由PUF蛋白识别靶mRNA的3′非翻译区内的特异性结合位点驱动。在这项研究中，我们研究了由创始PUF，果蝇Pumilio，和它的人类直系同源物的镇压机制。在这里，我们评估了先前提出的模型，其中Pumilio RNA结合结构域（RBD）结合Argonaute，这反过来又阻止了真核延伸因子1A的翻译活性。令人惊讶的是，我们发现Argonautes是不必要的抑制引起的果蝇和人类的PUFs在体内。第二个模型提出，Pumilio的RBD通过募集去腺苷酸酶来缩短mRNA的聚腺苷尾来抑制。事实上，RBD与Pop 2去腺苷酶结合并加速去腺苷化;然而，这种活性对于调节并不重要。相反，我们确定poly（A）是RBD抑制所必需的。我们的研究结果表明，多聚腺苷酸依赖性阻遏的RBD需要多聚腺苷酸结合蛋白，pAbp。此外，我们表明，由人RAB 2 RBD的镇压需要pAbp的直系同源物，PABPC 1。Pumilio与pAbp结合，但不破坏pAbp与mRNA的结合。总之，我们的数据支持一种模型，其中Pumilio RBD拮抗pAbp促进翻译的能力。因此，PUF RBD的保守功能是结合特异性mRNA，拮抗pAbp功能，并促进去腺苷化。
This article analyzes the mechanism by which Pumilio represses the translation of its targets. The results show, rather surprisingly, that promotion of deadenylation is not required for expression. Instead, Pumilio interacts with poly(A) binding protein and somehow interferes with its activity. PUF proteins are potent repressors that serve important roles in stem cell maintenance, neurological processes, and embryonic development. These functions are driven by PUF protein recognition of specific binding sites within the 3′ untranslated regions of target mRNAs. In this study, we investigated mechanisms of repression by the founding PUF, Drosophila Pumilio, and its human orthologs. Here, we evaluated a previously proposed model wherein the Pumilio RNA binding domain (RBD) binds Argonaute, which in turn blocks the translational activity of the eukaryotic elongation factor 1A. Surprisingly, we found that Argonautes are not necessary for repression elicited by Drosophila and human PUFs in vivo. A second model proposed that the RBD of Pumilio represses by recruiting deadenylases to shorten the mRNA's polyadenosine tail. Indeed, the RBD binds to the Pop2 deadenylase and accelerates deadenylation; however, this activity is not crucial for regulation. Rather, we determined that the poly(A) is necessary for repression by the RBD. Our results reveal that poly(A)-dependent repression by the RBD requires the poly(A) binding protein, pAbp. Furthermore, we show that repression by the human PUM2 RBD requires the pAbp ortholog, PABPC1. Pumilio associates with pAbp but does not disrupt binding of pAbp to the mRNA. Taken together, our data support a model wherein the Pumilio RBD antagonizes the ability of pAbp to promote translation. Thus, the conserved function of the PUF RBD is to bind specific mRNAs, antagonize pAbp function, and promote deadenylation.
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影响因子：
64.5
作者：
Hafner M;Landthaler M;Burger L;Khorshid M;Hausser J;Berninger P;Rothballer A;Ascano M Jr;Jungkamp AC;Munschauer M;Ulrich A;Wardle GS;Dewell S;Zavolan M;Tuschl T
通讯作者：
Tuschl T
DOI：
10.1042/bj20111474
发表时间：
2012-02-01
期刊：
The Biochemical journal
影响因子：
--
作者：
Brook M;McCracken L;Reddington JP;Lu ZL;Morrice NA;Gray NK
通讯作者：
Gray NK
DOI：
10.1073/pnas.201284398
发表时间：
2001-09-25
影响因子：
11.1
作者：
Chagnovich, D;Lehmann, R
通讯作者：
Lehmann, R
影响因子：
4.8
作者：
Hook, Brad A.;Goldstrohm, Aaron C.;Wickens, Marvin
通讯作者：
Wickens, Marvin
DOI：
10.1073/pnas.0603701103
发表时间：
2006-06-20
影响因子：
11.1
作者：
Karim, Muhammad M.;Svitkin, Yuri V.;Sonenberg, Nahum
通讯作者：
Sonenberg, Nahum