Requirements for the assembly and release of Newcastle disease virus-like particles
Graduate School of Biomedical Sciences; Program in Immunology and Virology; Department of Molecular Genetics and Microbiology
Medical Subject Headings
Animals; Cell Line; Cell Nucleus; Chickens; Cytoplasm; Electrophoresis, Polyacrylamide Gel; Immunoprecipitation; Microscopy, Confocal; Microscopy, Electron, Transmission; Models, Biological; Newcastle disease virus; Protein Interaction Mapping; Viral Structural Proteins; *Virus Assembly
Life Sciences | Medicine and Health Sciences
Paramyxoviruses, such as Newcastle disease virus (NDV), assemble in and bud from plasma membranes of infected cells. To explore the role of each of the NDV structural proteins in virion assembly and release, virus-like particles (VLPs) released from avian cells expressing all possible combinations of the nucleoprotein (NP), membrane or matrix protein (M), an uncleaved fusion protein (F-K115Q), and hemagglutinin-neuraminidase (HN) protein were characterized for densities, protein content, and efficiencies of release. Coexpression of all four proteins resulted in the release of VLPs with densities and efficiencies of release (1.18 to 1.16 g/cm(3) and 83.8% +/- 1.1%, respectively) similar to those of authentic virions. Expression of M protein alone, but not NP, F-K115Q, or HN protein individually, resulted in efficient VLP release, and expression of all different combinations of proteins in the absence of M protein did not result in particle release. Expression of any combination of proteins that included M protein yielded VLPs, although with different densities and efficiencies of release. To address the roles of NP, F, and HN proteins in VLP assembly, the interactions of proteins in VLPs formed with different combinations of viral proteins were characterized by coimmunoprecipitation. The colocalization of M protein with cell surface F and HN proteins in cells expressing all combinations of viral proteins was characterized. Taken together, the results show that M protein is necessary and sufficient for NDV budding. Furthermore, they suggest that M-HN and M-NP interactions are responsible for incorporation of HN and NP proteins into VLPs and that F protein is incorporated indirectly due to interactions with NP and HN protein.
Rights and Permissions
Citation: J Virol. 2006 Nov;80(22):11062-73. Epub 2006 Sep 13. Link to article on publisher's site
Journal of virology
Pantua, Homer Dadios; McGinnes, Lori; Peeples, Mark E.; and Morrison, Trudy G., "Requirements for the assembly and release of Newcastle disease virus-like particles" (2006). GSBS Student Publications. 931.