Masaki So, Yoshimoto Rei, Kaida Daisuke, Hata Asuka, Satoh Takayuki, Ohno Mutsuhito, Kataoka Naoyuki
Medical Innovation Center, Laboratory for Malignancy Control Research, Kyoto University Graduate School of Medicine, Sakyo-ku, Kyoto 606-8507, Japan.
Chemical Genetics Laboratory, RIKEN Advanced Science Institute, Wako, Saitama 351-0198, Japan.
Int J Mol Sci. 2015 Feb 9;16(2):3705-21. doi: 10.3390/ijms16023705.
In eukaryotes, pre-mRNA splicing is an essential step for gene expression. We have been analyzing post-splicing intron turnover steps in higher eukaryotes. Here, we report protein interaction between human Debranching enzyme 1 (hDbr1) and several factors found in the Intron Large (IL) complex, which is an intermediate complex of the intron degradation pathway. The hDbr1 protein specifically interacts with xeroderma pigmentosum, complementeation group A (XPA)-binding protein 2 (Xab2). We also attempted to identify specific interactors of hDbr1. Co-immunoprecipitation experiments followed by mass spectrometry analysis identified a novel protein as one of the specific interactors of hDbr1. This protein is well conserved among many species and shows the highest similarity to yeast Drn1, so it is designated as human Dbr1 associated ribonuclease 1 (hDrn1). hDrn1 directly interacts with hDbr1 through protein-protein interaction. Furthermore, hDrn1 shuttles between the nucleus and the cytoplasm, as hDbr1 protein does. These findings suggest that hDrn1 has roles in both the nucleus and the cytoplasm, which are highly likely to involve hDbr1.
在真核生物中,前体mRNA剪接是基因表达的一个关键步骤。我们一直在分析高等真核生物中剪接后内含子的周转步骤。在此,我们报告了人类去分支酶1(hDbr1)与在内含子大(IL)复合体中发现的几种因子之间的蛋白质相互作用,IL复合体是内含子降解途径的一个中间复合体。hDbr1蛋白特异性地与着色性干皮病A互补组(XPA)结合蛋白2(Xab2)相互作用。我们还试图鉴定hDbr1的特异性相互作用蛋白。通过免疫共沉淀实验结合质谱分析,鉴定出一种新蛋白作为hDbr1的特异性相互作用蛋白之一。这种蛋白在许多物种中高度保守,与酵母Drn1的相似性最高,因此被命名为人类Dbr1相关核糖核酸酶1(hDrn1)。hDrn1通过蛋白质-蛋白质相互作用直接与hDbr1相互作用。此外,hDrn1像hDbr1蛋白一样在细胞核和细胞质之间穿梭。这些发现表明hDrn1在细胞核和细胞质中都发挥作用,极有可能涉及hDbr1。