Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Japan.
Insect Mol Biol. 2012 Apr;21(2):247-56. doi: 10.1111/j.1365-2583.2011.01131.x. Epub 2012 Jan 9.
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.
从大蜡螟（Galleria mellonella）的中肠克隆并测序了编码半胱天冬酶-1（caspase-1）的 cDNA，caspase-1 是一种参与细胞凋亡的主要蛋白酶。开放阅读框包含 879 个核苷酸，编码 293 个氨基酸，并被注册为 Gmcaspase-1。序列比较显示与鳞翅目半胱天冬酶-1、人半胱天冬酶-3 和秀丽隐杆线虫的 ced-3 具有高度同源性。Gmcaspase-1 预计包含一个短的前导肽、大亚基和小亚基结构域。它还表现出半胱天冬酶的所有特征，包括在 Asp-25、Asp-192 和 Asp-181 后有三个保守的切割位点，三个活性位点残基包括高度保守的 QACQG 五肽活性位点基序，以及四个底物结合位点。发育过程中的表达谱表明，Gmcaspase-1 的转录本及其蛋白产物在变态过程中中肠的两个或更多波中出现。免疫组织化学、原位杂交和 TUNEL 分析表明，凋亡首先发生在中肠上皮的基底、中间和顶端区域，然后在腔中形成黄体。至少有三个波的有丝分裂和分化紧随凋亡波从基底和中间到顶端部分形成成年上皮。
