Cloning and functional characterizations of an apoptogenic Hid gene in the Scuttle Fly, Megaselia scalaris (Diptera; Phoridae).
Cloning and functional characterizations of an apoptogenic Hid gene in the Scuttle Fly, Megaselia scalaris (Diptera; Phoridae).
复制标题
Scuttle Fly, Megaselia scalaris（双翅目； Phoridae）中凋亡 Hid 基因的克隆和功能特征。
DOI：
10.1016/j.gene.2016.11.043
复制
发表时间：
2017
期刊：
影响因子：
3.5
通讯作者：
Park,JaeH
中科院分区：
文献类型：
--
作者：
Yoo,Siuk;Lam,Haylie;Lee,Chansong;Lee,Gyunghee;Park,JaeH
虽然果蝇的细胞凋亡死亡机制已经得到了很好的研究，但目前还不清楚这种机制是否在其他远亲物种中也是保守的。本研究利用简并引物和聚合酶链式反应(PCR)技术，克隆了烟粉虱Megaselia scalaris(MsHid)头部退化缺陷(HID)促凋亡基因，该基因编码197个氨基酸的多肽，是迄今为止最小的HID蛋白。PCR分析表明，MsHid基因由4个外显子和3个内含子组成。MsHidin在果蝇体内多种肽能神经元和非神经元组织中的异位表达能有效地诱导这些细胞的凋亡。然而，N端IBM或C端MTS保守结构域的缺失使MsHID的促凋亡活性丧失，表明这两个结构域是不可或缺的。MsHid在各生命阶段均有表达，但在胚胎和蛹中的表达更为显著，在中枢神经系统(CNS)中表达活跃，表明MsHid在中枢神经系统发育中起重要作用。总之，MsHID可能是该物种胚胎和胚胎后发育过程中重要的细胞死亡诱因。此外，我们还发现，在紫外线照射的胚胎中，MsHid的表达增加了2倍，这表明MsHidin在UV诱导的细胞凋亡中可能起到了作用。
Although the mechanisms of apoptotic cell death have been well studied in the fruit fly, Drosophila melanogaster, it is unclear whether such mechanisms are conserved in other distantly related species. Using degenerate primers and PCR, we cloned a proapoptotic gene homologous toHead involution defective(Hid) from the Scuttle fly,Megaselia scalaris(MsHid).MsHidcDNA encodes a 197-amino acid-long polypeptide, which so far is the smallest HID protein. PCR analyses revealed that theMsHidgene consists of four exons and three introns. Ectopic expression ofMsHidin various peptidergic neurons and non-neuronal tissues inDrosophilaeffectively induced apoptosis of these cells. However, deletion of either conserved domain, N-terminal IBM or C-terminal MTS, abolished the apoptogenic activity of MsHID, indicating that these two domains are indispensable. Expression ofMsHidwas found in all life stages, but more prominently in embryos and pupae.MsHidis actively expressed in the central nervous system (CNS), indicating its important role in CNS development. Together MsHID is likely to be an important cell death inducer during embryonic and post-embryonic development in this species. In addition, we found 2-fold induction ofMsHidexpression in UV-irradiated embryos, indicating a possible role forMsHidin UV-induced apoptosis.