Functional genomic analysis of the ADP-ribosylation factor family of GTPases:: phylogeny among diverse eukaryotes and function in C-elegans
Functional genomic analysis of the ADP-ribosylation factor family of GTPases:: phylogeny among diverse eukaryotes and function in C-elegans
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DOI：
10.1096/fj.04-2273com
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发表时间：
2004-12-01
期刊：
影响因子：
4.8
通讯作者：
Kahn, RA
中科院分区：
文献类型：
--
作者：
Li, YW;Kelly, WG;Kahn, RA
ADP核糖基化因子（Arf）和Arf样（Arl）蛋白是一类在真核生物进化早期就出现的高度保守的21 kDa GTP酶家族。这些蛋白质在囊泡运输、脂质代谢、微管动力学、发育和可能的其他细胞过程中起调节作用。我们发现的证据存在的6 Arf家族成员在原生动物蓝氏贾第鞭毛虫和22个成员在哺乳动物。进行系统发育分析，描绘Arf家族成员之间的进化关系，并试图组织他们的进化起源和功能，在细胞和/或生物体。从动物、真菌、植物和原生生物中分析的相似的100个蛋白质序列聚成11个组，包括Arfs、9个Arls和Sar蛋白。为了开始在后生动物模式生物中对该家族进行功能分析，我们研究了所有三种C。利用RNA介导的干扰（RNAi）技术，对线虫Arfs（Arf-1、Arf-3和Arf-6）和三种Arls（Arl-1、Arl-2和Arl-3）进行了克隆和测序。将编码Arf-1或Arf-3的双链RNA（dsRNA）注射到N2雌雄同体动物中，在其后代中产生胚胎致死性，随后在注射的动物本身中产生不育性。注射Arl-2 dsRNA导致早期后代的生殖系紊乱和不育，后期后代表现出早期胚胎停滞。因此，在C.线虫，胚胎发生至少需要三个。这些数据代表了对该家族多个成员在多细胞生物体发育中的作用的首次分析。
ADP-ribosylation factor (Arf) and Arf-like (Arl) proteins are a family of highly conserved 21 kDa GTPases that emerged early in the evolution of eukaryotes. These proteins serve regulatory roles in vesicular traffic, lipid metabolism, microtubule dynamics, development, and likely other cellular processes. We found evidence for the presence of 6 Arf family members in the protist Giardia lamblia and 22 members in mammals. A phylogenetic analysis was performed to delineate the evolutionary relationships among Arf family members and to attempt to organize them by both their evolutionary origins and functions in cells and/or organisms. The similar to 100 protein sequences analyzed from animals, fungi, plants, and protists clustered into 11 groups, including Arfs, nine Arls, and Sar proteins. To begin functional analyses of the family in a metazoan model organism, we examined roles for all three C. elegans Arfs (Arf-1, Arf-3, and Arf-6) and three Arls (Arl-1, Arl-2, and Arl-3) by use of RNA-mediated interference (RNAi). Injection of double-stranded RNA ( dsRNA) encoding Arf-1 or Arf-3 into N2 hermaphrodites produced embryonic lethality in their offspring and, later, sterility in the injected animals themselves. Injection of Arl-2 dsRNA resulted in a disorganized germline and sterility in early offspring, with later offspring exhibiting an early embryonic arrest. Thus, of the six Arf family members examined in C. elegans, at least three are required for embryogenesis. These data represent the first analysis of the role(s) of multiple members of this family in the development of a multicellular organism.