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Expression analyses of caspase‐1 and related activities in the midgut of Galleria mellonella during metamorphosis
Author(s) -
Khoa D. B.,
Trang L. T. D.,
Takeda M.
Publication year - 2012
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/j.1365-2583.2011.01131.x
Subject(s) - biology , midgut , microbiology and biotechnology , caenorhabditis elegans , open reading frame , galleria mellonella , complementary dna , peptide sequence , biochemistry , gene , botany , larva , virulence
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 Gm caspase‐1. The sequence comparison showed a high homology to lepidopteran caspase‐1, human caspase‐3, and ced‐3 of Caenorhabditis elegans . Gm caspase‐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 Gm caspase‐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.