Autographa californica Multiple Nucleopolyhedrovirus EXON0 (ORF141) Is Required for Efficient Egress of Nucleocapsids from the Nucleus
- 15 September 2007
- journal article
- Published by American Society for Microbiology in Journal of Virology
- Vol. 81 (18), 9859-9869
- https://doi.org/10.1128/jvi.00588-07
Abstract
Autographa californica multiple nucleopolyhedrovirus (AcMNPV) exon0 ( orf141 ) has been shown to be required for the efficient production of budded virus (BV). The deletion of exon0 reduces the level of BV production by up to 99% (X. Dai, T. M. Stewart, J. A. Pathakamuri, Q. Li, and D. A. Theilmann, J. Virol. 78:9633-9644, 2004); however, the function or mechanism by which EXON0 affects BV production is unknown. In this study, we further elucidated the function of EXON0 by investigating the localization of EXON0 in infected Sf9 cells and in virions and by identifying interactions between EXON0 and other viral proteins. In addition, electron microscopy was used to study the cellular localization of nucleocapsids in cells transfected with an exon0 knockout (KO) virus. The results showed that EXON0 was localized to both the cytoplasm and the nuclei of infected Sf9 cells throughout the infection. Western blotting results also showed that EXON0 was purified along with BV and occlusion-derived virus (ODV). The fractionation of BV into the nucleocapsid and envelope components showed that EXON0 localized to the BV nucleocapsid. Yeast two-hybrid screening, coimmunoprecipitation, and confocal microscopy revealed that it interacted with nucleocapsid proteins FP25 and BV/ODV-C42. Cells transfected with the exon0 KO virus exhibited normally appearing nucleocapsids in the nuclei in numbers equal to those in the nuclei of cells transfected with the EXON0 repaired virus. In contrast, the numbers of nucleocapsids in the cytoplasm of cells transfected with the exon0 KO virus were significantly lower than those in the cytoplasm of cells transfected with the repaired virus. These results support the conclusion that EXON0 is required in the BV pathway for the efficient egress of nucleocapsids from the nucleus to the cytoplasm.Keywords
This publication has 57 references indexed in Scilit:
- Autographa californica Multiple Nucleopolyhedrovirus Nucleocapsid Assembly Is Interrupted upon Deletion of the 38K GeneJournal of Virology, 2006
- Functional Entry of Baculovirus into Insect and Mammalian Cells Is Dependent on Clathrin-Mediated EndocytosisJournal of Virology, 2006
- The Immediate-Early Protein IE0 of the Autographa californica Nucleopolyhedrovirus Is Not Essential for Viral ReplicationJournal of Virology, 2005
- Herpes Simplex Virus Tegument Protein US11 Interacts with Conventional Kinesin Heavy ChainJournal of Virology, 2002
- Effects of Deletion and Overexpression of the Autographa californica Nuclear Polyhedrosis Virus FP25K Gene on Synthesis of Two Occlusion-Derived Virus Envelope Proteins and Their Transport into Virus-Induced Intranuclear MembranesJournal of Virology, 2001
- Acidic pH induces fusion of cells infected with baculovirus to form syncytiaFEBS Letters, 1992
- Ultrastructural observations of virion maturation in Autographa californica nuclear polyhedrosis virus infected Spodoptera frugiperda cell culturesJournal of Ultrastructure and Molecular Structure Research, 1986
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976
- Morphogenesis of nuclear polyhedrosis virus under conditions of prolonged passage in vitroJournal of Ultrastructure Research, 1974
- The Virogenic Stroma in Nuclear and Cytoplasmic PolyhedrosesNature, 1956