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Journal of Virology, March 2005, p. 2988-2997, Vol. 79, No. 5
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.5.2988-2997.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Evidence for a New Viral Late-Domain Core Sequence, FPIV, Necessary for Budding of a Paramyxovirus

Anthony P. Schmitt,1 George P. Leser,1 Eiji Morita,2 Wesley I. Sundquist,2 and Robert A. Lamb1,3*

Department of Biochemistry, Molecular Biology and Cell Biology,1 Howard Hughes Medical Institute, Northwestern University, Evanston, Illinois,3 Department of Biochemistry, University of Utah, Salt Lake City, Utah2

Received 7 September 2004/ Accepted 13 October 2004

Enveloped virus budding has been linked to both the ubiquitin-proteasome pathway and the vacuolar protein-sorting pathway of cells. We show here for the paramyxovirus SV5 that proteasome inhibitors and expression of dominant-negative VPS4(E228Q) ATPase blocks budding. The SV5 matrix (M) protein lacks previously defined late domains (e.g., P[T/S]AP, PPxY, YPDL) that recruit cellular factors. We identified a new motif for budding (core sequence FPIV) that can compensate functionally for lack of a PTAP late domain in budding human immunodeficiency virus type 1 virus-like particles (VLPs). Mutagenesis experiments suggest the more general sequence Ø-P-x-V. The proline residue was found to be critically important for function of this sequence, as substitution of this proline in the SV5 M protein resulted in poor budding of SV5 VLPs and failure of recombinant SV5 virus to replicate normally. Adaptation of mutant virus occurred rapidly, resulting in new proline residues elsewhere in the M protein. We hypothesize that these proline residues act to partially restore virus budding by generation of new motifs that act as suboptimal late domains.

* Corresponding author. Mailing address: BMBCB, Northwestern University, 2205 Tech Dr., Evanston, IL 60208-3500. Phone: (847) 491-5433. Fax: (847) 491-2467. E-mail: ralamb{at}northwestern.edu.

Journal of Virology, March 2005, p. 2988-2997, Vol. 79, No. 5
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.5.2988-2997.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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