Shen Qing-Ji, Kassim Hakimi, Huang Yong, Li Hui, Zhang Jing, Li Guang, Wang Peng-Ye, Yan Jun, Ye Fangfu, Liu Ji-Long
Medical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT, UK.
Medical Research Council Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT, UK; Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400715, China.
J Genet Genomics. 2016 Jun 20;43(6):393-404. doi: 10.1016/j.jgg.2016.03.008. Epub 2016 Apr 1.
Compartmentation via filamentation has recently emerged as a novel mechanism for metabolic regulation. In order to identify filament-forming metabolic enzymes systematically, we performed a genome-wide screening of all strains available from an open reading frame-GFP collection in Saccharomyces cerevisiae. We discovered nine novel filament-forming proteins and also confirmed those identified previously. From the 4159 strains, we found 23 proteins, mostly metabolic enzymes, which are capable of forming filaments in vivo. In silico protein-protein interaction analysis suggests that these filament-forming proteins can be clustered into several groups, including translational initiation machinery and glucose and nitrogen metabolic pathways. Using glutamine-utilising enzymes as examples, we found that the culture conditions affect the occurrence and length of the metabolic filaments. Furthermore, we found that two CTP synthases (Ura7p and Ura8p) and two asparagine synthetases (Asn1p and Asn2p) form filaments both in the cytoplasm and in the nucleus. Live imaging analyses suggest that metabolic filaments undergo sub-diffusion. Taken together, our genome-wide screening identifies additional filament-forming proteins in S. cerevisiae and suggests that filamentation of metabolic enzymes is more general than currently appreciated.
通过丝状化进行区室化最近已成为一种新的代谢调节机制。为了系统地鉴定形成丝状的代谢酶,我们对酿酒酵母开放阅读框-GFP文库中所有可用菌株进行了全基因组筛选。我们发现了9种新的形成丝状的蛋白质,并证实了之前鉴定出的那些。在4159个菌株中,我们发现了23种蛋白质,其中大部分是代谢酶,它们能够在体内形成丝状。计算机模拟的蛋白质-蛋白质相互作用分析表明,这些形成丝状的蛋白质可以聚类为几个组,包括翻译起始机制以及葡萄糖和氮代谢途径。以谷氨酰胺利用酶为例,我们发现培养条件会影响代谢丝的出现和长度。此外,我们发现两种CTP合酶(Ura7p和Ura8p)以及两种天冬酰胺合成酶(Asn1p和Asn2p)在细胞质和细胞核中均形成丝状。实时成像分析表明代谢丝经历亚扩散。综上所述,我们的全基因组筛选鉴定出了酿酒酵母中更多的形成丝状的蛋白质,并表明代谢酶的丝状化比目前所认识到的更为普遍。