Müller F, Krüger D, Sattlegger E, Hoffmann B, Ballario P, Kanaan M, Barthelmess I B
Institut für Angewandte Genetik, Universität Hannover, Germany.
Mol Gen Genet. 1995 Jul 28;248(2):162-73. doi: 10.1007/BF02190797.
Phenotypic and molecular studies of the mutation U142 indicate that the cpc-2+ gene is required to activate general amino acid control under conditions of amino acid limitation in the vegetative growth phase, and for formation of protoperithecia in preparation for the sexual phase of the life cycle of Neurospora crassa. The cpc-2 gene was cloned by complementation of the cpc-2 mutation in a his-2ts bradytrophic background. Genomic and cDNA sequence analysis indicated a 1636 bp long open reading frame interrupted by four introns. The deduced 316 amino acid polypeptide reveals 70% positional identity over its full length with G-protein beta-subunit-related polypeptides found in humans, rat (RACK1), chicken, tobacco and Chlamydomonas. With the exception of RACK1 the function of these proteins is obscure. All are entirely made up of seven WD-repeats. Expression studies of cpc-2 revealed one abundant transcript in the wild type; in the mutant its level is drastically reduced. In mutant cells transformed with the complementing sequence, the transcript level, enzyme regulation and female fertility are restored. In the wild type the cpc-2 transcript is down-regulated under conditions of amino acid limitation. With cpc-2 a new element involved in general amino acid control has been identified, indicating a function for a WD-repeat protein that belongs to a class that is conserved throughout the evolution of eukaryotes.
对突变体U142的表型和分子研究表明,cpc-2+基因在营养生长阶段氨基酸限制条件下激活一般氨基酸控制是必需的,并且对于粗糙脉孢菌生命周期有性阶段准备过程中原雌器的形成也是必需的。通过在his-2ts生长缓慢营养缺陷型背景下对cpc-2突变体进行互补克隆了cpc-2基因。基因组和cDNA序列分析表明,一个1636 bp长的开放阅读框被四个内含子打断。推导的316个氨基酸的多肽在其全长上与在人类、大鼠(RACK1)、鸡、烟草和衣藻中发现的G蛋白β亚基相关多肽有70%的位置同一性。除RACK1外,这些蛋白质的功能尚不清楚。它们全部由七个WD重复序列组成。cpc-2的表达研究表明,野生型中有一个丰富的转录本;在突变体中其水平急剧降低。在用互补序列转化的突变体细胞中,转录本水平、酶调节和雌性育性得以恢复。在野生型中,cpc-2转录本在氨基酸限制条件下被下调。通过cpc-2鉴定出了一个参与一般氨基酸控制的新元件,表明一种WD重复蛋白具有一种在真核生物进化过程中保守的功能。