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https://doi.org/10.1186/1743-422X-10-294
Title: | Evidence for the interaction of the human metapneumovirus G and F proteins during virus-like particle formation | Authors: | Loo, L.H Jumat, M.R Fu, Y Ayi, T.C Wong, P.S Tee, N.W.S Tan, B.H Sugrue, R.J |
Keywords: | Human metapneumovirus f protein Human metapneumovirus g protein Human metapneumovirus m protein unclassified drug virus protein article electron microscopy human human cell Human metapneumovirus image analysis mammal cell nonhuman nucleotide sequence protein cross linking protein expression protein protein interaction sucrose density gradient centrifugation sucrose gradient virus assembly virus like agent virus morphology Human metapneumovirus Mammalia Cell Line Centrifugation, Density Gradient Child Child, Preschool Gene Expression Glycoproteins Humans Metapneumovirus Microscopy, Electron Molecular Sequence Data Protein Binding Recombinant Proteins RNA, Viral Sequence Analysis, DNA Viral Fusion Proteins Viral Proteins Virosomes |
Issue Date: | 2013 | Citation: | Loo, L.H, Jumat, M.R, Fu, Y, Ayi, T.C, Wong, P.S, Tee, N.W.S, Tan, B.H, Sugrue, R.J (2013). Evidence for the interaction of the human metapneumovirus G and F proteins during virus-like particle formation. Virology Journal 10 : 294. ScholarBank@NUS Repository. https://doi.org/10.1186/1743-422X-10-294 | Rights: | Attribution 4.0 International | Abstract: | Background: Human metapneumovirus (HMPV) is now a major cause of lower respiratory infection in children. Although primary isolation of HMPV has been achieved in several different cell lines, the low level of virus replication and the subsequent recovery of low levels of infectious HMPV have hampered biochemical studies on the virus. These experimental methodologies usually require higher levels of biological material that can be achieved following HMPV infection. In this study we demonstrate that expression of the HMPV F, G and M proteins in mammalian cells leads to HMPV virus-like particles (VLP) formation. This experimental strategy will serve as a model system to allow the process of HMPV virus assembly to be examined. Methods. The HMPV F, G and M proteins were expressed in mammalian cell lines. Protein cross-linking studies, sucrose gradient centrifugation and in situ imaging was used to examine interactions between the virus proteins. VLP formation was examined using sucrose density gradient centrifugation and electron microscopy analysis. Results: Analysis of cells co-expressing the F, G and M proteins demonstrated that these proteins interacted. Furthermore, in cells co-expression the three HMPV proteins the formation VLPs was observed. Image analysis revealed the VLPs had a similar morphology to the filamentous virus morphology that we observed on HMPV-infected cells. The capacity of each protein to initiate VLP formation was examined using a VLP formation assay. Individual expression of each virus protein showed that the G protein was able to form VLPs in the absence of the other virus proteins. Furthermore, co-expression of the G protein with either the M or F proteins facilitated their incorporation into the VLP fraction. Conclusion: Co-expression of the F, G and M proteins leads to the formation of VLPs, and that incorporation of the F and M proteins into VLPs is facilitated by their interaction with the G protein. Our data suggests that the G protein plays a central role in VLP formation, and further suggests that the G protein may also play a role in the recruitment of the F and M proteins to sites of virus particle formation during HMPV infection. © 2013 Loo et al.; licensee BioMed Central Ltd. | Source Title: | Virology Journal | URI: | https://scholarbank.nus.edu.sg/handle/10635/181801 | ISSN: | 1743422X | DOI: | 10.1186/1743-422X-10-294 | Rights: | Attribution 4.0 International |
Appears in Collections: | Staff Publications Elements |
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