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Journal of Virology, October 2007, p. 11363-11371, Vol. 81, No. 20
0022-538X/07/$08.00+0     doi:10.1128/JVI.01281-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Pseudorabies Virus Us9 Directs Axonal Sorting of Viral Capsids{triangledown} ,{dagger}

M. G. Lyman,1 B. Feierbach,1,2 D. Curanovic,1 M. Bisher,1 and L. W. Enquist1*

Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544,1 Lewis-Sigler Institute for Integrative Genomics, Princeton, New Jersey 085442

Received 12 June 2007/ Accepted 31 July 2007

Pseudorabies virus (PRV) mutants lacking the Us9 gene cannot spread from presynaptic to postsynaptic neurons in the rat visual system, although retrograde spread remains unaffected. We sought to recapitulate these findings in vitro using the isolator chamber system developed in our lab for analysis of the transneuronal spread of infection. The wild-type PRV Becker strain spreads efficiently to postsynaptic neurons in vitro, whereas the Us9-null strain does not. As determined by indirect immunofluorescence, the axons of Us9-null infected neurons do not contain the glycoproteins gB and gE, suggesting that their axonal sorting is dependent on Us9. Importantly, we failed to detect viral capsids in the axons of Us9-null infected neurons. We confirmed this observation by using three different techniques: by direct fluorescence of green fluorescent protein-tagged capsids; by transmission electron microscopy; and by live-cell imaging in cultured, sympathetic neurons. This finding has broad impact on two competing models for how virus particles are trafficked inside axons during anterograde transport and redefines a role for Us9 in viral sorting and transport.

* Corresponding author. Mailing address: 314 Schultz Laboratory, Department of Molecular Biology, Princeton University, Princeton, NJ 08544. Phone: (609) 258-2415. Fax: (609) 258-1035. E-mail: lenquist{at}princeton.edu

{triangledown} Published ahead of print on 8 August 2007.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Virology, October 2007, p. 11363-11371, Vol. 81, No. 20
0022-538X/07/$08.00+0     doi:10.1128/JVI.01281-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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