Structure of the N-terminal Mlp1-binding domain of the Saccharomyces cerevisiae mRNA-binding protein, Nab2.

Structure of the N-terminal Mlp1-binding domain of the Saccharomyces cerevisiae mRNA-binding protein, Nab2.
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DOI:
10.1016/j.jmb.2007.11.087
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发表时间:
2008-02-29
影响因子:
5.6
通讯作者:
Stewart, Murray
Stewart, Murray
中科院分区:
生物学2区
文献类型:
--
作者:
Grant, Richard P.;Marshall, Neil J.;Yang, Ji-Chun;Fasken, Milo B.;Kelly, Seth M.;Harrernan, Michelle T.;Neuhaus, David;Corbett, Anita H.;Stewart, Murray

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核丰富的多聚腺苷酸RNA结合蛋白2(Nab 2)是一种重要的酵母核异质性核糖核蛋白,它调节mRNA核输出和多聚腺苷酸尾长。Nab 2的N-末端结构域(残基1-97)介导与核孔相关蛋白、肌球蛋白样蛋白1(Mlp 1)的C-末端球状结构域和mRNA输出因子Gfd 1的相互作用。Nab 2 N-末端结构域的溶液和晶体结构显示出主要螺旋折叠,其类似于在其他几种RNA结合蛋白中发现的PWI折叠。与其他含PWI的蛋白质相比,我们没有发现Nab 2 N-末端结构域与核酸结合的证据。相反,这个结构域似乎介导蛋白质:蛋白质相互作用,促进mRNA的核输出。Nab 2 N-末端结构域具有以Phe 73为中心的独特疏水补丁,与该表面区域是蛋白质:蛋白质相互作用位点一致。在这个疏水补丁内的工程突变减弱了与Mlp 1 C-末端结构域的相互作用,但不改变与Gfd 1的相互作用,表明该补丁形成Nab 2和Mlp 1之间的界面的关键组成部分。
Nuclear abundant poly(A) RNA-binding protein 2 (Nab2) is an essential yeast heterogeneous nuclear ribonucleoprotein that modulates both mRNA nuclear export and poly(A) tail length. The N-terminal domain of Nab2 (residues 1–97) mediates interactions with both the C-terminal globular domain of the nuclear pore-associated protein, myosin-like protein 1 (Mlp1), and the mRNA export factor, Gfd1. The solution and crystal structures of the Nab2 N-terminal domain show a primarily helical fold that is analogous to the PWI fold found in several other RNA-binding proteins. In contrast to other PWI-containing proteins, we find no evidence that the Nab2 N-terminal domain binds to nucleic acids. Instead, this domain appears to mediate protein:protein interactions that facilitate the nuclear export of mRNA. The Nab2 N-terminal domain has a distinctive hydrophobic patch centered on Phe73, consistent with this region of the surface being a protein:protein interaction site. Engineered mutations within this hydrophobic patch attenuate the interaction with the Mlp1 C-terminal domain but do not alter the interaction with Gfd1, indicating that this patch forms a crucial component of the interface between Nab2 and Mlp1.
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