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酵母RNA结合蛋白Rbp1p可改变线粒体孔蛋白mRNA的稳定性。

The yeast RNA-binding protein Rbp1p modifies the stability of mitochondrial porin mRNA.

作者信息

Buu Leh-Miauh, Jang Li-Ting, Lee Fang-Jen S

机构信息

Institute of Molecular Medicine, College of Medicine, National Taiwan University Hospital, National Taiwan University, 7 Chung Shan South Road, Taipei 100, Taiwan.

出版信息

J Biol Chem. 2004 Jan 2;279(1):453-62. doi: 10.1074/jbc.M309278200. Epub 2003 Oct 21.

Abstract

The Saccharomyces cerevisiae RNA-binding protein Rbp1p was initially identified as a negative growth regulator; however, its function is still obscure. Here, we show that Rbp1p in cells is associated with structures that sediment at 10,000 as well as 100,000 x g. It appears microscopically as punctate signals partially localized to the perinuclear region. Over-expression of Rbp1p in yeast resulted in growth defects on nonfermentable carbon sources, suggesting a function for Rbp1p in mitochondrial biogenesis. Absence of Rbp1p increased the level of mitochondrial porin, whereas over-expression of Rbp1p, but not an N-terminally truncated form, decreased porin levels. Over-expression of Rbp1p also decreased the level of mitochondrial porin mRNA by enhancing its degradation, an effect that was dependent on all three of the Rbp1p RNA recognition motifs. In cells, the porin mRNA is associated with Rbp1p.RNP (ribonucleoprotein) complexes. In vitro binding assays showed that Rbp1p most likely interacts with a (C/G)U-rich element in the porin mRNA 3'-UTR. Based on these observations, we infer that Rbp1p has a role in negatively regulating mitochondrial porin expression post-transcriptionally.

摘要

酿酒酵母RNA结合蛋白Rbp1p最初被鉴定为一种负生长调节因子;然而,其功能仍不清楚。在这里,我们表明细胞中的Rbp1p与在10000倍重力以及100000倍重力下沉淀的结构相关联。在显微镜下,它表现为部分定位于核周区域的点状信号。在酵母中过表达Rbp1p会导致在非发酵碳源上生长缺陷,这表明Rbp1p在线粒体生物发生中具有功能。缺失Rbp1p会增加线粒体孔蛋白的水平,而过表达Rbp1p(但不是N端截短形式)会降低孔蛋白水平。过表达Rbp1p还通过增强其降解来降低线粒体孔蛋白mRNA的水平,这种效应依赖于Rbp1p的所有三个RNA识别基序。在细胞中,孔蛋白mRNA与Rbp1p.RNP(核糖核蛋白)复合物相关联。体外结合试验表明,Rbp1p很可能与孔蛋白mRNA 3'-UTR中富含(C/G)U的元件相互作用。基于这些观察结果,我们推断Rbp1p在转录后对线粒体孔蛋白表达起负调节作用。

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