Murdoch Kirstie, Loop Susanne, Rudt Falko, Pieler Tomas
Institut für Biochemie und Molekulare Zellbiologie, Georg-August-Universität, Göttingen, Germany.
Eur J Cell Biol. 2002 Oct;81(10):549-56. doi: 10.1078/0171-9335-00271.
In Xenopus oocytes, 5S rRNA is exported out of the nucleus in the context of two ribonucleoprotein complexes (RNPs): complexed with transcription factor IIIA as the 7S RNP or as the 5S RNP with ribosomal protein L5. 5S rRNA-containing RNP export takes place at a slow rate in comparison to that of nuclear export signal-containing proteins and the U1 snRNP. Using oocyte microinjection assays we found that the export of 5S RNPs requires nuclear RanGTP and RanGTP hydrolysis and is leptomycin B-sensitive, indicating the process is mediated by the export receptor CRM1. A novel nuclear export signal motif is characterised in a region of L5 also possessing a nuclear import signal, thus identifying a shuttling domain for this protein. This same motif in L5 is found to be required for interaction with CRM1 in vitro and for export in vivo.
在非洲爪蟾卵母细胞中,5S核糖体RNA(rRNA)在两种核糖核蛋白复合体(RNP)的背景下被输出细胞核:与转录因子IIIA结合形成7S RNP,或与核糖体蛋白L5结合形成5S RNP。与含核输出信号的蛋白质和U1小核核糖核蛋白(snRNP)相比,含5S rRNA的RNP输出速率较慢。通过卵母细胞显微注射实验,我们发现5S RNP的输出需要细胞核中的RanGTP及其水解,并且对雷帕霉素B敏感,这表明该过程由输出受体CRM1介导。一个新的核输出信号基序在L5的一个区域被鉴定出来,该区域也具有核输入信号,从而确定了该蛋白的一个穿梭结构域。发现L5中的这个相同基序在体外与CRM1相互作用以及在体内输出过程中都是必需的。