RAE1 is a shuttling mRNA export factor that binds to a GLEBS-like NUP98 motif at the nuclear pore complex through multiple domains.
RAE1 is a shuttling mRNA export factor that binds to a GLEBS-like NUP98 motif at the nuclear pore complex through multiple domains.
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DOI：
10.1083/jcb.145.2.237
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发表时间：
1999-04-19
影响因子：
7.8
通讯作者：
van Deursen, J M
中科院分区：
文献类型：
--
作者：
Pritchard, C E;Fornerod, M;Kasper, L H;van Deursen, J M
Gle 2 p参与酿酒酵母中poly（A）+ RNA的核输出和核孔复合物（NPC）的结构和分布。Gle 2 p通过Nup 116 p内的短Gle 2 p结合基序（称为GLEBS）锚定在核膜（NE）上。Gle 2 p和Gle 2 p-Nup 116 p相互作用在mRNA输出中起作用的分子机制尚不清楚。在这里，我们表明，RAE 1，Gle 2 p的哺乳动物同源物，结合GLEBS样NUP 98基序在NPC通过多个域，包括WD重复和COOH末端非WD重复延伸。这种相互作用是直接的，如体外结合研究和化学交联所证明的。在非洲爪蟾卵母细胞中进行的显微注射实验表明，RAE 1穿梭于细胞核和细胞质之间，并以温度依赖性和RanGTP独立的方式从细胞核中输出。RAE 1与NE的对接高度依赖于新mRNA的合成。GLEBS样基序的过表达也抑制NE与RAE 1的结合并诱导poly（A）+ RNA的核积累。这两种效应都被废除，通过在GLEBS样基序中引入点突变或通过RAE 1的过表达，表明RAE 1和NUP 98-RAE 1相互作用在mRNA输出中的直接作用。总之，我们的数据表明，RAE 1是一种穿梭运输因子，通过其锚在NPC的特定NUP 98基序的能力，直接有助于mRNA的核输出。
Gle2p is implicated in nuclear export of poly(A)+ RNA and nuclear pore complex (NPC) structure and distribution in Saccharomyces cerevisiae. Gle2p is anchored at the nuclear envelope (NE) via a short Gle2p-binding motif within Nup116p called GLEBS. The molecular mechanism by which Gle2p and the Gle2p–Nup116p interaction function in mRNA export is unknown. Here we show that RAE1, the mammalian homologue of Gle2p, binds to a GLEBS-like NUP98 motif at the NPC through multiple domains that include WD-repeats and a COOH-terminal non–WD-repeat extension. This interaction is direct, as evidenced by in vitro binding studies and chemical cross-linking. Microinjection experiments performed in Xenopus laevis oocytes demonstrate that RAE1 shuttles between the nucleus and the cytoplasm and is exported from the nucleus in a temperature-dependent and RanGTP-independent manner. Docking of RAE1 to the NE is highly dependent on new mRNA synthesis. Overexpression of the GLEBS-like motif also inhibits NE binding of RAE1 and induces nuclear accumulation of poly(A)+ RNA. Both effects are abrogated either by the introduction of point mutations in the GLEBS-like motif or by overexpression of RAE1, indicating a direct role for RAE1 and the NUP98–RAE1 interaction in mRNA export. Together, our data suggest that RAE1 is a shuttling transport factor that directly contributes to nuclear export of mRNAs through its ability to anchor to a specific NUP98 motif at the NPC.