Investigating the consequences of eIF4E2 (4EHP) interaction with 4E-transporter on its cellular distribution in HeLa cells.
Investigating the consequences of eIF4E2 (4EHP) interaction with 4E-transporter on its cellular distribution in HeLa cells.
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DOI：
10.1371/journal.pone.0072761
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Standart N
中科院分区：
文献类型：
--
作者：
Kubacka D;Kamenska A;Broomhead H;Minshall N;Darzynkiewicz E;Standart N
除了典型的eIF 4 E帽结合蛋白之外，真核生物还进化出具有不同特征的序列相关变体，其中一些变体已被证明负调控特定mRNA的翻译，但其特征仍然很差。哺乳动物eIF 4 E蛋白分为三类，第一类代表典型的帽结合蛋白eIF 4 E1。eIF 4 E1结合eIF 4G以启动翻译，其他eIF 4 E结合蛋白如4 E-BP和4 E-T通过将eIF 4 E1与相同的共有序列YX 4L β结合来阻止这种相互作用。我们研究了人eIF 4 E2（4 EHP）与eIF 4 E转运蛋白4 E-T的相互作用。我们首先发现eIF 4 E1：4 E-T的比例在HeLa和HEK 293细胞中分别为50：1至15：1，而eIF 4 E2：4 E-T的比例在6：1至3：1之间变化。我们接下来提供的证据表明，eIF 4 E2结合4 E-T在酵母双杂交测定，以及在下拉测定和招募到哺乳动物细胞中的P-体。我们还表明，虽然eIF 4 E1和eIF 4 E2通过典型的YX 4L β序列结合4 E-T，附近的下游序列也影响eIF 4 E：4 E-T相互作用。间接免疫荧光被用来证明，eIF 4 E2，通常均匀地定位在细胞质中，不重新分配到应力颗粒亚砷酸盐处理的细胞，也不是P-体放线菌素D处理的细胞，在对比eIF 4 E1。此外，eIF 4 E2以Crm 1依赖的方式穿梭通过细胞核，但以4 E-T独立的方式，也不同于eIF 4 E1。总之，我们得出结论，虽然这两个帽结合蛋白与4 E-T相互作用，并可以招募4 E-T的P体，eIF 4 E2的功能可能是不同的eIF 4 E1，无论是在细胞质和细胞核，进一步扩展我们的理解哺乳动物I类和II类帽结合蛋白。
In addition to the canonical eIF4E cap-binding protein, eukaryotes have evolved sequence–related variants with distinct features, some of which have been shown to negatively regulate translation of particular mRNAs, but which remain poorly characterised. Mammalian eIF4E proteins have been divided into three classes, with class I representing the canonical cap-binding protein eIF4E1. eIF4E1 binds eIF4G to initiate translation, and other eIF4E-binding proteins such as 4E-BPs and 4E-T prevent this interaction by binding eIF4E1 with the same consensus sequence YX 4Lϕ. We investigate here the interaction of human eIF4E2 (4EHP), a class II eIF4E protein, which binds the cap weakly, with eIF4E-transporter protein, 4E-T. We first show that ratios of eIF4E1:4E-T range from 50:1 to 15:1 in HeLa and HEK293 cells respectively, while those of eIF4E2:4E-T vary from 6:1 to 3:1. We next provide evidence that eIF4E2 binds 4E-T in the yeast two hybrid assay, as well as in pull-down assays and by recruitment to P-bodies in mammalian cells. We also show that while both eIF4E1 and eIF4E2 bind 4E-T via the canonical YX 4Lϕ sequence, nearby downstream sequences also influence eIF4E:4E-T interactions. Indirect immunofluorescence was used to demonstrate that eIF4E2, normally homogeneously localised in the cytoplasm, does not redistribute to stress granules in arsenite-treated cells, nor to P-bodies in Actinomycin D-treated cells, in contrast to eIF4E1. Moreover, eIF4E2 shuttles through nuclei in a Crm1-dependent manner, but in an 4E-T–independent manner, also unlike eIF4E1. Altogether we conclude that while both cap-binding proteins interact with 4E-T, and can be recruited by 4E-T to P-bodies, eIF4E2 functions are likely to be distinct from those of eIF4E1, both in the cytoplasm and nucleus, further extending our understanding of mammalian class I and II cap-binding proteins.
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影响因子：
5.3
作者：
Cargnello, Marie;Tcherkezian, Joseph;Roux, Philippe P.
通讯作者：
Roux, Philippe P.
影响因子：
8.8
作者：
Culjkovic-Kraljacic B;Baguet A;Volpon L;Amri A;Borden KL
通讯作者：
Borden KL
影响因子：
3.4
作者：
Joshi, B;Lee, K;Maeder, DL;Jagus, R
通讯作者：
Jagus, R
DOI：
10.1073/pnas.1005320107
发表时间：
2010-08-10
影响因子：
11.1
作者：
Furic, Luc;Rong, Liwei;Sonenberg, Nahum
通讯作者：
Sonenberg, Nahum
DOI：
10.1083/jcb.200504039
发表时间：
2005-09-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ferraiuolo MA;Basak S;Dostie J;Murray EL;Schoenberg DR;Sonenberg N
通讯作者：
Sonenberg N