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Journal of Virology, January 2006, p. 460-473, Vol. 80, No. 1
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.1.460-473.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Nonstructural Protein 3 of Bluetongue Virus Assists Virus Release by Recruiting ESCRT-I Protein Tsg101

Christoph Wirblich, Bishnupriya Bhattacharya, and Polly Roy*

Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London WC1E 7HT, United Kingdom

Received 2 June 2005/ Accepted 29 September 2005

The release of Bluetongue virus (BTV) and other members of the Orbivirus genus from infected host cells occurs predominantly by cell lysis, and in some cases, by budding from the plasma membrane. Two nonstructural proteins, NS3 and NS3A, have been implicated in this process. Here we show that both proteins bind to human Tsg101 and its ortholog from Drosophila melanogaster with similar strengths in vitro. This interaction is mediated by a conserved PSAP motif in NS3 and appears to play a role in virus release. The depletion of Tsg101 with small interfering RNA inhibits the release of BTV and African horse sickness virus, a related orbivirus, from HeLa cells up to fivefold and threefold, respectively. Like most other viral proteins which recruit Tsg101, NS3 also harbors a PPXY late-domain motif that allows NS3 to bind NEDD4-like ubiquitin ligases in vitro. However, the late-domain motifs in NS3 do not function as effectively in facilitating the release of mini Gag virus-like particles from 293T cells as the late domains from human immunodeficiency virus type 1, human T-cell leukemia virus, and Ebola virus. A mutagenesis study showed that the arginine residue in the PPRY motif is responsible for the low activity of the NS3 late-domain motifs. Our data suggest that the BTV late-domain motifs either recruit an antagonist that interferes with budding or fail to recruit an agonist which is different from NEDD4.

* Corresponding author. Mailing address: London School of Hygiene and Tropical Medicine, Keppel St., London WC1E 7HT, United Kingdom. Phone: 44 020-79272324. Fax: 44 020-79272839. E-mail: polly.roy{at}lshtm.ac.uk.

Journal of Virology, January 2006, p. 460-473, Vol. 80, No. 1
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.1.460-473.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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