Protein-protein interactions between the testis brain RNA-binding protein and the transitional endoplasmic reticulum ATPase, a cytoskeletal gamma actin and Trax in male germ cells and the brain.
Protein-protein interactions between the testis brain RNA-binding protein and the transitional endoplasmic reticulum ATPase, a cytoskeletal gamma actin and Trax in male germ cells and the brain.
复制标题
睾丸脑 RNA 结合蛋白与移行内质网 ATP 酶、男性生殖细胞和大脑中的细胞骨架伽玛肌动蛋白和 Trax 之间的蛋白质-蛋白质相互作用。
DOI：
10.1021/bi990573s
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Hecht,NB
中科院分区：
文献类型：
--
作者：
Wu,XQ;Lefrancois,S;Morales,CR;Hecht,NB
睾丸脑RNA结合蛋白（TB-RBP）及其人类同源物翻译蛋白（Translin）具有多种功能，从mRNA转运和翻译调节到DNA重排和修复。为了深入了解这种26 kDa蛋白质的可能功能，免疫沉淀法用于鉴定小鼠胞质提取物中与TB-RBP相互作用的蛋白质。三种蛋白质，即过渡型内质网ATP酶、细胞骨架γ肌动蛋白和Trax，与重组小鼠TB-RBP的亲和纯化抗体特异性免疫沉淀。重组蛋白的体外结合试验和EM免疫细胞化学证实了TB-RBP在体外和体内与TER ATP酶相互作用。共聚焦显微镜已经证明TB-RBP与肌动蛋白共定位在雄性生殖细胞的细胞质中。Trax与TB-RBP的免疫沉淀证实了已发表的报告，该报告证明了酵母双杂交测定中两种蛋白质之间的蛋白质相互作用。这些数据支持TB-RBP作为将特定mRNA连接到细胞骨架结构的连接物的假设，并表明广泛表达的TER ATP酶参与细胞内和/或细胞间mRNA转运。
Numerous functions have been proposed for the testis brain RNA-binding protein (TB-RBP) and its human homologue, Translin, ranging from mRNA transport and translational regulation to DNA rearrangement and repair. To gain insight into the likely functions of this 26 kDa protein, immunoprecipitation was used to identify proteins that interact with TB-RBP in mouse cytosolic extracts. Three proteins, the transitional endoplasmic reticulum ATPase, a cytoskeletal γ actin, and Trax, were specifically immunoprecipitated with an affinity-purified antibody to recombinant mouse TB-RBP. In vitro binding assays with recombinant proteins and EM immunocytochemistry confirm that TB-RBP interacts with the TER ATPase in vitro and in vivo. Confocal microscopy has demonstrated that TB-RBP colocalizes with actin in the cytoplasm of male germ cells. The immunoprecipitation of Trax with TB-RBP confirms a published report demonstrating protein interactions between the two proteins in a yeast two-hybrid assay. These data support the hypothesis that TB-RBP serves as a link in attaching specific mRNAs to cytoskeletal structures and suggests an involvement for the ubiquitously expressed TER ATPase in intracellular and/or intercellular mRNA transport.