Hahm B, Cho O H, Kim J E, Kim Y K, Kim J H, Oh Y L, Jang S K
Department of Life Science, Pohang University of Science and Technology, Kyungbuk, South Korea.
FEBS Lett. 1998 Apr 3;425(3):401-6. doi: 10.1016/s0014-5793(98)00269-5.
Polypyrimidine tract-binding protein (PTB) is involved in pre-mRNA splicing and internal ribosomal entry site (IRES)-dependent translation. In order to identify cellular protein(s) interacting with PTB, we performed a yeast two-hybrid screening. Heterogeneous nuclear ribonucleoprotein L (hnRNP L) was identified as a PTB-binding protein. The interaction between PTB and hnRNP L was confirmed in an in vitro binding assay. Both PTB and hnRNP L were found to localize in the nucleoplasm, excepting the nucleoli, in HeLa cells by the green fluorescent protein (GFP)-fused protein detection method. The N-terminal half of PTB (aa 1-329) and most of hnRNP L (aa 141-558) is required for the interaction between PTB and hnRNP L.
聚嘧啶序列结合蛋白（PTB）参与前体mRNA剪接和内部核糖体进入位点（IRES）依赖性翻译。为了鉴定与PTB相互作用的细胞蛋白，我们进行了酵母双杂交筛选。异质性核核糖核蛋白L（hnRNP L）被鉴定为PTB结合蛋白。通过体外结合试验证实了PTB与hnRNP L之间的相互作用。通过绿色荧光蛋白（GFP）融合蛋白检测方法发现，在HeLa细胞中，PTB和hnRNP L均定位于核质中，核仁除外。PTB与hnRNP L之间的相互作用需要PTB的N端一半（氨基酸1 - 329）和hnRNP L的大部分（氨基酸141 - 558）。
