SfDronc,an initiator caspase involved in apoptosis in the fall armyworm Spodoptera frugiperda |
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| Authors: | Ning Huang Srgjan Civciristov Christine J Hawkins Rollie J Clem |
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| Institution: | 1. Molecular, Cellular and Developmental Biology Program, Division of Biology, Kansas State University, Manhattan, KS 66503, USA;2. Department of Biochemistry, La Trobe University, Bundoora 3086, Victoria, Australia;1. Department of Biology, The Pennsylvania State University, 208 Mueller Laboratory, University Park, PA 16802, USA;2. Department of Plant Science, The Pennsylvania State University, University Park, PA 16802, USA;3. United States Department of Agriculture USDA-ARS, Purdue University, West Lafayette, IN, USA;4. Department of Plant and Soil Sciences, The AgBase Databases, Institute for Genomics, Biocomputing and Biotechnology (IGBB), HPC2, Box 9555, Mississippi State University, MS 39762, USA;1. Department of Entomology, Louisiana State University Agricultural Center, 404 Life Sciences Building, Baton Rouge, LA 70803, USA;2. Department of Entomology and Nematology, University of Florida, IFAS SWFREC, Immokalee, FL 34142, USA;1. Departamento de Entomologia, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil;2. Instituto Nacional de Ciência e Tecnologia em Interações Planta-Praga, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil;3. DuPont Pioneer, 7100 NW 62nd Avenue, Johnston, IA, 50131, United States;1. INRA, UMR 1355, Institut Sophia Agrobiotech, 400 Route des Chappes, 06903 Sophia-Antipolis, France;2. CNRS, UMR 7254, 400 Route des Chappes, 06903 Sophia-Antipolis, France;3. Université de Nice Sophia Antipolis, 400 Route des Chappes, 06903 Sophia-Antipolis, France;4. Environnement Canada, Centre Saint-Laurent, 105 Mc Gill Street, Montreal, QC, Canada H2Y 2E7 |
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| Abstract: | Initiator caspases are the first caspases that are activated following an apoptotic stimulus, and are responsible for cleaving and activating downstream effector caspases, which directly cause apoptosis. We have cloned a cDNA encoding an ortholog of the initiator caspase Dronc in the lepidopteran insect Spodoptera frugiperda. The SfDronc cDNA encodes a predicted protein of 447 amino acids with a molecular weight of 51 kDa. Overexpression of SfDronc induced apoptosis in Sf9 cells, while partial silencing of SfDronc expression in Sf9 cells reduced apoptosis induced by baculovirus infection or by treatment with UV or actinomycin D. Recombinant SfDronc exhibited several expected biochemical characteristics of an apoptotic initiator caspase: 1) SfDronc efficiently cleaved synthetic initiator caspase substrates, but had very little activity against effector caspase substrates; 2) mutation of a predicted cleavage site at position D340 blocked autoprocessing of recombinant SfDronc and reduced enzyme activity by approximately 10-fold; 3) SfDronc cleaved the effector caspase Sf-caspase-1 at the expected cleavage site, resulting in Sf-caspase-1 activation; and 4) SfDronc was strongly inhibited by the baculovirus caspase inhibitor SpliP49, but not by the related protein AcP35. These results indicate that SfDronc is an initiator caspase involved in caspase-dependent apoptosis in S. frugiperda, and as such is likely to be responsible for the initiator caspase activity in S. frugiperda cells known as Sf-caspase-X. |
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