Molecular alliance of Lymantria dispar multiple nucleopolyhedrovirus and a short unmodified antisense oligonucleotide of its anti-apoptotic IAP-3 gene: A novel approach for gypsy moth control

Baculovirus IAP (inhibitor-of-apoptosis) genes originated by capture of host genes. Unmodified short antisense DNA oligonucleotides (oligoDNAs) from baculovirus IAP genes can down-regulate specific gene expression profiles in both baculovirus-free and baculovirus-infected insects. In this study, gypsy moth (Lymantria dispar) larvae infected with multiple nucleopolyhedrovirus (LdMNPV), and LdMNPV-free larvae, were treated with oligoDNA antisense to the RING (really interesting new gene) domain of the LdMNPV IAP-3 gene. The results with respect to insect mortality, biomass accumulation, histological studies, RT-PCR, and analysis of DNA apoptotic fragmentation suggest that oligoRING induced increased apoptotic processes in both LdMNPV-free and LdMNPV-infected insect cells, but were more pronounced in the latter. These data open up possibilities for promising new routes of insect pest control using antisense phosphodiester DNA oligonucleotides

Molecular Alliance of Lymantria dispar Multiple Nucleopolyhedrovirus and a Short Unmodified Antisense Oligonucleotide of Its Anti-Apoptotic IAP-3 Gene: A Novel Approach for Gypsy Moth Control

Baculovirus IAP (inhibitor-of-apoptosis) genes originated by capture of host genes. Unmodified short antisense DNA oligonucleotides (oligoDNAs) from baculovirus IAP genes can down-regulate specific gene expression profiles in both baculovirus-free and baculovirus-infected insects. In this study, gypsy moth (Lymantria dispar) larvae infected with multiple nucleopolyhedrovirus (LdMNPV), and LdMNPV-free larvae, were treated with oligoDNA antisense to the RING (really interesting new gene) domain of the LdMNPV IAP-3 gene. The results with respect to insect mortality, biomass accumulation, histological studies, RT-PCR, and analysis of DNA apoptotic fragmentation suggest that oligoRING induced increased apoptotic processes in both LdMNPV-free and LdMNPV-infected insect cells, but were more pronounced in the latter. These data open up possibilities for promising new routes of insect pest control using antisense phosphodiester DNA oligonucleotides

Beet yellows virus replicase and replicative compartments: parallels with other RNA viruses

In eukaryotic virus systems, infection leads to induction of membranous compartments in which replication occurs. Virus-encoded subunits of the replication complex mediate its interaction with membranes. As replication platforms, RNA viruses use the cytoplasmic surfaces of different membrane compartments, e.g., endoplasmic reticulum (ER), Golgi, endo/lysosomes, mitochondria, chloroplasts and peroxisomes. Closterovirus infections are accompanied by formation of multivesicular complexes from cell membranes of ER or mitochondrial origin. So far the mechanisms for vesicles formation have been obscure. In the replication-associated 1a polyprotein of Beet yellows virus (BYV) and other closteroviruses, the region between the methyltransferase (MTR) and helicase (HEL) domains (1a central region, 1a CR) is marginally conserved. Computer-assisted analysis predicts several putative membrane-binding domains in the BYV 1a CR. Transient expression of a hydrophobic segment (referred to here as CR-2) of the BYV 1a in Nicotiana benthamiana led to reorganization of the ER and formation of ~1-m mobile globules. We propose that the CR-2 may be involved in the formation of multivesicular complexes in BYV-infected cells. This provides analogy with membrane-associated proteins mediating the build-up of virus factories in cells infected with diverse positive-strand RNA viruses (alpha-like viruses, picorna-like viruses, flaviviruses, and nidoviruses) and negative-strand RNA viruses (bunyaviruses)

Topical treatment of LdMNPV-infected gypsy moth caterpillars with 18 nucleotides long antisense fragment from LdMNPV IAP3 gene triggers higher levels of apoptosis in infected cells and mortality of the pest

The high efficiency of baculovirus infection is partially explained by the ability of the virus to suppress host defense machinery connected with the apoptosis pathway. Members of the baculovirus gene family, inhibitors of apoptosis (vIAPs), have been shown to inhibit apoptosis in baculovirus-infected cells. Here we showed that treatment of the LdMNPV- -infected 1st instar gypsy moth (Lymantria dispar) caterpillars with sense (oligoBIR) and antisense (oligoRING) DNA oligonucleotides from the LdMNPV IAP3 gene induced elevated mortality of the insects. Apoptotic DNA ladder assay showed that the leading role in this phenomenon is played by the antisense oligoRING fragment of the vIAP3 gene. These results imply that the application of both antisense DNA oligonucleotides from vIAP genes and baculovirus preparations (one following the other) may be a potential method for plant protection against insect pests

Original Article. Topical treatment of LdMNPV-infected gypsy moth caterpillars with 18 nucleotides long antisense fragment from LdMNPV IAP3 gene triggers higher levels of apoptosis in infected cells and mortality of the pest

The high efficiency of baculovirus infection is partially explained by the ability of the virus to suppress host defense machinery connected with the apoptosis pathway. Members of the baculovirus gene family, inhibitors of apoptosis (vIAPs), have been shown to inhibit apoptosis in baculovirus-infected cells. Here we showed that treatment of the LdMNPV- -infected 1st instar gypsy moth (Lymantria dispar) caterpillars with sense (oligoBIR) and antisense (oligoRING) DNA oligonucleotides from the LdMNPV IAP3 gene induced elevated mortality of the insects. Apoptotic DNA ladder assay showed that the leading role in this phenomenon is played by the antisense oligoRING fragment of the vIAP3 gene. These results imply that the application of both antisense DNA oligonucleotides from vIAP genes and baculovirus preparations (one following the other) may be a potential method for plant protection against insect pests