■翻译是生活的基本过程。在真核生物中,翻译的延伸步骤是高度保守的,并且由真核翻译延伸因子(eEF)1A和eEF2驱动。延伸的一个显着变化是酿酒酵母中真核延伸因子(eEF)3的活性,由酵母延伸因子(YEF3)与所有真菌物种中的直向同源物编码,一些藻类,还有一些原生生物.在酿酒酵母中,YEF3是必需基因,eEF3在翻译延伸中起关键作用,因为它促进三元复合物酰化转移RNA(tRNA)-eEF1A-鸟苷-5'-三磷酸(GTP)与核糖体的氨酰基(A)位点的结合,肽易位后不带电的tRNA的释放,和核糖体回收。尽管YEF3在40多年前被发现,eEF3几乎只在酿酒酵母中表征。
■我们进行了一种体内遗传学方法,以评估eEF3在系统发育遥远的真菌物种中的功能保守性。
■我们发现来自鲁氏酵母和光滑假丝酵母(均属于子囊门)的eEF3,Ustilagomaydis(担子菌门),和性腺增生(单藻门),但不是构巢曲霉(子囊门),支持缺乏内源性YEF3基因的酿酒酵母的生长。我们还证明了eEF3是子囊菌C.glabrata和A.nidulans中的必需基因。
■鉴于大多数关于真菌翻译的现有知识仅来自酿酒酵母,我们在该生物体以外的发现显示了真菌伸长过程的变异性。我们还证明了eEF3在病原真菌中至关重要,开启了使用eEF3作为目标对抗念珠菌病的可能性。
UNASSIGNED: Translation is a fundamental process of life. In eukaryotes, the elongation step of translation is highly conserved and is driven by eukaryotic translation elongation factors (eEF)1A and eEF2. A significant variation of the elongation is the activity of eukaryotic elongation factor (eEF) 3 in Saccharomyces cerevisiae encoded by the gene yeast elongation factor (YEF3) with orthologs in all fungal species, a few algae, and some protists. In S. cerevisiae, YEF3 is an essential gene and eEF3 plays a critical role in translation elongation, as it promotes binding of the ternary complex acylated-Transfer RNA (tRNA)-eEF1A-Guanosine-5\'-triphosphate (GTP) to the aminoacyl (A) site of the ribosome, the release of uncharged tRNAs after peptide translocation, and ribosome recycling. Even though YEF3 was discovered more than 40 years ago, eEF3 has been characterized almost exclusively in S. cerevisiae.
UNASSIGNED: We undertook an in vivo genetic approach to assess the functional conservation of eEF3 across phylogenetically distant fungal species.
UNASSIGNED: We found that eEF3 from Zygosaccharomyces rouxii and Candida glabrata (both belonging to phylum Ascomycota), Ustilago maydis (phylum Basidiomycota), and Gonapodya prolifera (phylum Monoblepharomycota), but not Aspergillus nidulans (phylum Ascomycota), supported the growth of S. cerevisiae lacking the endogenous YEF3 gene. We also proved that eEF3 is an essential gene in the ascomycetes C. glabrata and A. nidulans.
UNASSIGNED: Given that most existing knowledge on fungal translation has only been obtained from S. cerevisiae, our findings beyond this organism showed variability in the elongation process in Fungi. We also proved that eEF3 is essential in pathogenic fungi, opening the possibility of using eEF3 as a target to fight candidiasis.