Biomaterial Database

Silencing of the baculovirus Op-iap3 gene by RNA interference reveals that it is required for prevention of apoptosis during Orgyia pseudotsugata M nucleopolyhedrovirus infection of Ld652Y cells. 2003

John C Means, and Israel Muro, and Rollie J Clem

Molecular, Cellular, and Developmental Biology Program, Division of Biology, Kansas State University, Manhattan, Kansas 66506, USA.

The Op-iap3 gene from the baculovirus Orgyia pseudotsugata M nucleopolyhedrovirus (OpMNPV) inhibits apoptosis induced by a mutant of Autographa californica MNPV (AcMNPV) that lacks the antiapoptotic gene p35, as well as apoptosis induced by a wide range of other stimuli in both mammalian and insect cells. However, the role of Op-iap3 during OpMNPV infection has not been previously examined. To determine the function of the Op-IAP3 protein during OpMNPV infection, we used RNA interference (RNAi) to silence Op-iap3 expression during OpMNPV infection of Ld652Y cells. Infected cells treated with Op-iap3 double-stranded RNA (dsRNA) did not accumulate detectable Op-iap3 mRNA, confirming that the Op-iap3 gene was effectively silenced. Op-IAP3 protein was found to be a component of the budded virion; however, in OpMNPV-infected cells treated with Op-iap3 dsRNA, the Op-IAP3 protein that was introduced by the inoculum virus decreased to almost undetectable levels by 12 h after dsRNA addition. Apoptosis was observed in infected cells treated with Op-iap3 dsRNA beginning at 12 h, and by 48 h, almost all of the cells had undergone apoptosis. These results show for the first time that Op-IAP3 is necessary to prevent apoptosis during OpMNPV infection. In addition, our results demonstrate that the RNAi technique can be an effective tool for studying baculovirus gene function.

UI	MeSH Term	Description	Entries
D007915	Lepidoptera	A large order of insects comprising the butterflies and moths.
D002478	Cells, Cultured	Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.	Cultured Cells,Cell, Cultured,Cultured Cell
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D014764	Viral Proteins	Proteins found in any species of virus.	Gene Products, Viral,Viral Gene Products,Viral Gene Proteins,Viral Protein,Protein, Viral,Proteins, Viral
D016367	Baculoviridae	Family of INSECT VIRUSES which contain polyhedron-shaped or ovocylindrical occlusion bodies. The genera include ALPHABACULOVIRUS; GAMMABACULOVIRUS; and DELTABACULOVIRUS (commonly known as NUCLEOPOLYHEDROVIRUSES) and BETABACULOVIRUS (commonly known as GRANULOVIRUSES). Baculovirus vectors are used for expression of foreign genes in insects and as BIOPESTICIDES for controlling insect populations.	Baculovirus,Baculoviruses
D017209	Apoptosis	A regulated cell death mechanism characterized by distinctive morphologic changes in the nucleus and cytoplasm, including the endonucleolytic cleavage of genomic DNA, at regularly spaced, internucleosomal sites, i.e., DNA FRAGMENTATION. It is genetically programmed and serves as a balance to mitosis in regulating the size of animal tissues and in mediating pathologic processes associated with tumor growth.	Apoptosis, Extrinsic Pathway,Apoptosis, Intrinsic Pathway,Caspase-Dependent Apoptosis,Classic Apoptosis,Classical Apoptosis,Programmed Cell Death,Programmed Cell Death, Type I,Apoptoses, Extrinsic Pathway,Apoptoses, Intrinsic Pathway,Apoptosis, Caspase-Dependent,Apoptosis, Classic,Apoptosis, Classical,Caspase Dependent Apoptosis,Cell Death, Programmed,Classic Apoptoses,Extrinsic Pathway Apoptoses,Extrinsic Pathway Apoptosis,Intrinsic Pathway Apoptoses,Intrinsic Pathway Apoptosis
D051034	Inhibitor of Apoptosis Proteins	A conserved class of proteins that control APOPTOSIS in both VERTEBRATES and INVERTEBRATES. IAP proteins interact with and inhibit CASPASES, and they function as ANTI-APOPTOTIC PROTEINS. The protein class is defined by an approximately 80-amino acid motif called the baculoviral inhibitor of apoptosis repeat; some members have ubiquitin-protein ligase activity.	BIRC2 Protein,Baculoviral IAP Repeat Containing Protein 2,Baculoviral IAP Repeat-Containing 2 Protein,Baculoviral Inhibitor Of Apoptosis Repeat Proteins,Cellular Inhibitor of Apoptosis 1 Protein,IAP Protein (Apoptosis),IAP Repeat Containing Proteins,IAP2 Protein,Inhibitor of Apoptosis 2 Protein,Inhibitor-Of-Apoptosis Protein,c-IAP1 Protein,cIAP1 Protein,Apoptosis Proteins Inhibitor,Baculoviral IAP Repeat Containing 2 Protein,Inhibitor Of Apoptosis Protein
D017924	Nucleopolyhedroviruses	Genera of the family BACULOVIRIDAE, characterized by the formation of crystalline, polyhedral occlusion bodies in the host cell nucleus. Nucleopolyhedrovirus is the common name for viruses from the Alphabaculovirus, Deltabaculovirus, or Gammabaculovirus genera.	Deltabaculovirus,Nuclear Polyhedrosis Virus,Nucleopolyhedrovirus,Alphabaculovirus,Gammabaculovirus,Alphabaculoviruses,Deltabaculoviruses,Gammabaculoviruses,Nuclear Polyhedrosis Viruses,Polyhedrosis Virus, Nuclear,Polyhedrosis Viruses, Nuclear,Virus, Nuclear Polyhedrosis,Viruses, Nuclear Polyhedrosis
D018411	Spodoptera	A genus of owlet moths of the family Noctuidae. These insects are used in molecular biology studies during all stages of their life cycle.	Spodoptera frugiperda,Spodoptera frugiperdas,Spodopteras,frugiperdas, Spodoptera
D020868	Gene Silencing	Interruption or suppression of the expression of a gene at transcriptional or translational levels.	Gene Inactivation,Inactivation, Gene,Silencing, Gene