Characterization of the biochemical properties of the human Upf1 gene product that is involved in nonsense-mediated mRNA decay
Characterization of the biochemical properties of the human Upf1 gene product that is involved in nonsense-mediated mRNA decay
复制标题
DOI：
10.1017/s1355838200000546
复制
发表时间：
2000-09-01
期刊：
影响因子：
4.5
通讯作者：
Peltz, SW
中科院分区：
文献类型：
--
作者：
Bhattacharya, A;Czaplinski, K;Peltz, SW
酵母中的Upf1蛋白参与了有效翻译终止的调节以及含有过早终止密码子的mRNA的加速周转，这种现象称为无义介导的mRNA衰变（NMD）。酵母UPF1的人类同源物，称为HUpf1/RENT1，也已被确定。HUpf1也被证明在哺乳动物细胞的NMD中发挥作用。酵母和人类UPF1蛋白的比较表明，氨基酸端半胱氨酸/组氨酸丰富区域和包含将该蛋白定义为超家族I族解旋酶的结构域的区域是保守的。酵母Upf1p显示rna依赖性atp酶和5‘ - >3 ’解旋酶活性。在本文中，我们报道了人类Upf1蛋白的表达、纯化和活性表征。我们证明了人类Upf1蛋白具有核酸依赖性atp酶活性和5‘ - bbbb3 ’解旋酶活性。此外，人类Upf1是一种rna结合蛋白，其rna结合活性由ATP调节。综上所述，这些结果表明Upf1蛋白的活性在物种间是保守的，反映了该蛋白在整个进化过程中的功能保守性。
The Upf1 protein in yeast has been implicated in the modulation of efficient translation termination as well as in the accelerated turnover of mRNAs containing premature stop codons, a phenomenon called nonsense-mediated mRNA decay (NMD). A human homolog of the yeast UPF1, termed HUpf1/RENT1, has also been identified. The HUpf1 has also been shown to play a role in NMD in mammalian cells. Comparison of the yeast and human UPF1 proteins demonstrated that the amino terminal cysteine/histidine-rich region and the region comprising the domains that define this protein as a superfamily group I helicase have been conserved. The yeast Upf1p demonstrates RNA-dependent ATPase and 5' --> 3' helicase activities. In this paper, we report the expression, purification, and characterization of the activities of the human Upf1 protein. We demonstrate that human Upf1 protein displays a nucleic-acid-dependent ATPase activity and a 5' --> 3' helicase activity. Furthermore, human Upf1 is an RNA-binding protein whose RNA-binding activity is modulated by ATP. Taken together, these results Indicate that the activities of the Upf1 protein are conserved across species, reflecting the conservation of function of this protein throughout evolution.