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Journal of Virology, December 2007, p. 13435-13443, Vol. 81, No. 24
0022-538X/07/$08.00+0     doi:10.1128/JVI.01469-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Intracellular Processing, Glycosylation, and Cell Surface Expression of Human Metapneumovirus Attachment Glycoprotein{triangledown}

Li Liu,1 Nathalie Bastien,2 and Yan Li1,2*

Department of Medical Microbiology and Infectious Diseases, the University of Manitoba, Winnipeg, Manitoba, Canada,1 National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada2

Received 5 July 2007/ Accepted 25 September 2007

The biosynthesis and posttranslational processing of human metapneumovirus attachment G glycoprotein were investigated. After pulse-labeling, the G protein accumulated as three species with molecular weights of 45,000, 50,000, and 53,000 (45K, 50K, and 53K, respectively). N-Glycosidase digestion indicated that these forms represent the unglycosylated precursor and N-glycosylated intermediate products, respectively. After an appropriate chase, these three naive forms were further processed to a mature 97K form. The presence of O-linked sugars in mature G protein was confirmed by O-glycanase digestion and lectin-binding assay using Arachis hypogaea (peanut agglutinin), an O-glycan-specific lectin. In addition, in the O-glycosylation-deficient cell line (CHO ldlD cell), the G protein could not be processed to the mature form unless the exogenous Gal and GalNAc were supplemented, which provided added evidence supporting the O-linked glycosylation of G protein. The maturation of G was completely blocked by monensin but was partially sensitive to brefeldin A (BFA), suggesting the O-linked glycosylation of G initiated in the trans-Golgi compartment and terminated in the trans-Golgi network. Enzymatic deglycosylation analysis confirmed that the BFA-G was a partial mature form containing N-linked oligosaccharides and various amounts of O-linked carbohydrate side chains. The expression of G protein at the cell surface could be detected by indirect immunofluorescence staining assay. Furthermore, cell surface immunoprecipitation displayed an efficient intracellular transport of G protein.

* Corresponding author. Mailing address: Influenza and Respiratory Viruses Section, National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba R3E 3R2, Canada. Phone: (204) 789-6045. Fax: (204) 789-2082. E-mail: yan_li{at}phac-aspc.gc.ca

{triangledown} Published ahead of print on 3 October 2007.

Journal of Virology, December 2007, p. 13435-13443, Vol. 81, No. 24
0022-538X/07/$08.00+0     doi:10.1128/JVI.01469-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Thammawat, S., Sadlon, T. A., Hallsworth, P. G., Gordon, D. L. (2008). Role of Cellular Glycosaminoglycans and Charged Regions of Viral G Protein in Human Metapneumovirus Infection. J. Virol. 82: 11767-11774 [Abstract] [Full Text]  


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