Expression analyses of caspase-1 and related activities in the midgut of Galleria mellonella during metamorphosis
Expression analyses of caspase-1 and related activities in the midgut of Galleria mellonella during metamorphosis
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DOI：
10.1111/j.1365-2583.2011.01131.x
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发表时间：
2012-04-01
影响因子：
2.6
通讯作者：
Takeda, M.
中科院分区：
文献类型：
--
作者：
Khoa, D. B.;Trang, L. T. D.;Takeda, M.
编码 caspase-1（一种参与细胞凋亡的主要蛋白酶）的 cDNA 是从大蜡蛾（Galleria melonella）的中肠中克隆并测序的。该开放阅读框含有879个核苷酸，编码293个氨基酸，注册为Gmcaspase-1。序列比对显示与鳞翅目caspase-1、人caspase-3、秀丽隐杆线虫ced-3具有高度同源性。 Gmcaspase-1 预计包含一个短前结构域、大亚基和小亚基结构域。它还表现出 caspase 的所有特征，包括 Asp-25、Asp-192 和 Asp-181 之后的三个保守切割位点、三个活性位点残基（包括高度保守的 QACQG 五肽活性位点基序）和四个底物结合位点。发育过程中的表达谱表明，Gmcaspase-1 及其蛋白产物的转录本在变态过程中在中肠中以两个或多个波出现。免疫组织化学、原位杂交和TUNEL分析显示，中肠上皮细胞的细胞凋亡首先发生在基底区，然后是中部区，然后是顶端区，并且在管腔中形成黄体。细胞凋亡波之后，从基底、中部到顶端至少经历三波有丝分裂和分化，形成成体上皮。
The cDNA encoding caspase-1, a main protease involved in apoptosis, was cloned and sequenced from the midgut of the greater wax moth, Galleria mellonella. The open reading frame contains 879 nucleotides, encodes 293 amino acids, and was registered as Gmcaspase-1. The sequence comparison showed a high homology to lepidopteran caspase-1, human caspase-3, and ced-3 of Caenorhabditis elegans. Gmcaspase-1 is predicted to contain a short prodomain, large subunit, and small subunit domain. It also exhibits all characteristics of caspase, including three conserved cleavage sites after Asp-25, Asp-192, and Asp-181, three active site residues including a highly conserved QACQG pentapeptide active-site motif, and four substrate binding sites. The expression profiles during development showed that the transcript of Gmcaspase-1 and its protein products appeared in two or more waves in the midgut during metamorphosis. Immunohistochemistry, in situ hybridization, and TUNEL analyses revealed that apoptosis occurred first at the basal, then middle and then apical regions in the midgut epithelium and the yellow body is formed in the lumen. At least three waves of mitosis and differentiation follow the apoptosis waves from the basal and middle to apical parts to form the adult epithelium.