Functional analysis of inhibitor of apoptosis 1 of the silkworm Bombyx mori. (December 2016)

• Record Type: Journal Article
• Authors: Hamajima, Rina; Iwamoto, Asako; Tomizaki, Moe; Suganuma, Ikue; Kitaguchi, Koji; Kobayashi, Michihiro; Yamada, Hayato; Ikeda, Motoko
• Is Part Of: Insect biochemistry and molecular biology. Volume 79(2016:Dec.)
• Page Start: 97
• Page End: 107
• Publication Date: 2016-12
• Subjects: Inhibitor of apoptosis -- IAP -- Apoptosis -- Bombyx mori -- Initiator caspase Dronc -- Lepidopteran caspase-1; Insect biochemistry -- Periodicals; Insects -- Physiology -- Periodicals; Insects -- Molecular aspects -- Periodicals; Biochemistry -- Periodicals; Insectes -- Biochimie -- Périodiques; Insectes -- Composition -- Périodiques; Insectes -- Physiologie -- Périodiques; Insectes -- Aspect moléculaire -- Périodiques; Biochimie -- Périodiques; Biochemistry; Insect biochemistry; Insects -- Molecular aspects; Insects -- Physiology; Periodicals; 572.8157
• DOI: 10.1016/j.ibmb.2016.10.012
• Languages: English
• ISSNs: 0965-1748

Abstract

Abstract: Recent advances in genome-wide surveys have revealed a number of lepidopteran insect homologs of mammalian and Drosophila genes that are responsible for apoptosis regulation. However, the underlying molecular mechanisms for apoptosis regulation in lepidopteran insect cells remain poorly understood. In the present study, we demonstrated that the transfection of Bombyx mori BM-N cells with dsRNA against the B. mori cellular iap1 gene ( cbm-iap1 ) induces severe apoptosis that is accompanied by an increase of caspase-3-like protease activity. In these apoptotic cells, the cleaved form of the endogenous initiator caspase Dronc (Bm-Dronc) was detected, indicating that cBm-IAP1 protein depletion by RNAi silencing resulted in the activation of Bm-Dronc. In transient expression assays in BM-N cells, cBm-IAP1 suppressed the apoptosis triggered by Bm-Dronc overexpression and depressed the elevation of caspase-3-like protease activity, but also increased the cleaved form of Bm-Dronc protein. cBm-IAP1 also suppressed the caspase-3-like protease activity stimulated by Bm-caspase-1 overexpression. Co-immunoprecipitation experiments demonstrated that cBm-IAP1 strongly interacts with Bm-Dronc, but only has weak affinity for Bm-caspase-1. Transient expression analyses showed that truncated cBm-IAP1 proteins defective in the BIR1, BIR2 or RING domain were unable to suppress Bm-Dronc-induced apoptosis. In addition, BM-N cells expressing truncated cBm-IAP1 proteins underwent apoptosis, suggesting that intact cBm-IAP1, which has anti-apoptotic activity, was replaced or displaced by the overexpressed truncated cBm-IAP1 proteins, which are incapable of interfering with the apoptotic caspase cascade. Taken together, the present results demonstrate that cBm-IAP1 is a vital negative regulator of apoptosis in BM-N cells and functions by preventing the activation and/or activity of Bm-Dronc and Bm-caspase-1. Graphical abstract: Highlights: Bombyx mori IAP (cBm-IAP1) depletion by RNAi-mediated silencing induced apoptosis in BM-N cells. cBm-IAP1 suppressed the apoptosis triggered by transient expression of the initiator caspase Bm-Dronc. cBm-IAP1 interacted strongly with Bm-Dronc, but only weakly with the effector caspase Bm-caspase-1. cBm-IAP1 protein defective in the BIR or RING finger region induced apoptosis in BM-N cells.