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Journal of Virology, December 2008, p. 11767-11774, Vol. 82, No. 23
0022-538X/08/$08.00+0     doi:10.1128/JVI.01208-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Role of Cellular Glycosaminoglycans and Charged Regions of Viral G Protein in Human Metapneumovirus Infection{triangledown}

Sutthiwan Thammawat, Tania A. Sadlon, Peter G. Hallsworth, and David L. Gordon*

Department of Microbiology and Infectious Diseases, Flinders University and Flinders Medical Centre, Bedford Park, Adelaide, Australia

Received 11 June 2008/ Accepted 2 September 2008

Human metapneumovirus (hMPV) is an important cause of lower respiratory tract disease, particularly in infants and young children. hMPV has two major glycoproteins, G and F, which are responsible for virus attachment and membrane fusion, respectively. We investigated the role of cellular glycosaminoglycans (GAGs) and G protein in hMPV infection. The pretreatment of hMPV with soluble heparin markedly inhibited the infection of HEp-2 cells. Recombinant G protein, comprising the extracellular domain of G, bound to heparin-agarose columns and also to HEp-2 cells. hMPV infection and G protein binding to HEp-2 cells was inhibited by other soluble GAGs, including chondroitin sulfates, by the enzymatic removal of cell surface GAGs with GAG lyases or by the pretreatment of cells with basic fibroblast growth factor. The role of cellular GAGs was confirmed by the binding of G protein to wild-type CHO cells but not to GAG-deficient CHO-pgsA745 cells. An analysis of the G protein sequence revealed two adjacent clusters of positively charged amino acids (149EKKKTRA155 and 159QRRGKGKE166). Truncated G fragments were expressed, and only the fragment containing these putative heparin binding domains retained heparin binding. The alanine mutagenesis of charged residues in either of these regions resulted in the loss of binding to heparin and to HEp-2 cells, suggesting that both sites are likely to be required for hMPV attachment. These results, taken together with the inhibition of hMPV infection by soluble G protein, indicate an important role for G protein and cellular GAGs in hMPV infection.

* Corresponding author. Mailing address: Department of Microbiology and Infectious Diseases, Flinders Medical Centre, Bedford Park, SA 5042, Australia. Phone: 61 8 82045293. Fax: 61 8 82044773. E-mail: D.Gordon{at}flinders.edu.au

{triangledown} Published ahead of print on 10 September 2008.

Journal of Virology, December 2008, p. 11767-11774, Vol. 82, No. 23
0022-538X/08/$08.00+0     doi:10.1128/JVI.01208-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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