Supporting Information

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Movie 1. Cotransport of capsids and VP1/2 after entry into neurons. mRFP1 capsid and GFP-VP1/2 images are presented as a color overlay. Six particles move through an axon toward the cell body during the recording (the axon, although not visible, crosses the field from bottom left to top right). Four additional particles remain static and are likely extracellular virions (three of the particles are not in contact with the axon).

Movie 2. Egress transport of capsids and VP16. mRFP1 capsid (Top), VP16-GFP (Middle), and a color merge (Bottom) are shown. The cell body from which this axon extends is beyond the left edge of the field of view. One egressing capsid is seen moving anterograde in the axon and is associated with VP16-GFP. Several additional VP16-GFP punctae of varying intensity move anterograde, independent of capsids.

Fig. 7. Description of fluorescent viruses. (A) Illustration of the PRV genome with characteristic internal and terminal repeats indicated (IR and TR, respectively). The region of the genome relevant to this report is expanded to show coding sequences, including those targeted for fusion with egfp (tegument-encoding genes UL36, UL37, UL47, UL48, and UL49) and mRFP1 (capsid-encoding gene UL35). Coding regions are scaled proportionately, and a summary of the PRV-expressed fusion proteins is shown below. Each dual-fluorescent virus encodes mRFP1-VP26 and a single GFP tegument fusion. (B) Growth kinetics of fluorescent and nonfluorescent viruses. Virions were harvested from the media (dashed lines, open symbols) and cells (solid lines, filled symbols) at indicated times. The mRFP1 capsid monofluorescent virus (PRV-GS847) was included in each experiment to allow for comparison between the graphs.

Fig. 8. Model of virus axonal transport. All tegument proteins are structural components of infecting virions, but some are selectively removed upon entering an axon. During egress, several tegument proteins transport by a capsid-independent mechanism to axon terminals, and their association with capsids is coincident with capsid targeting to the axon terminal. The illustration reflects the previous finding that VP 1/2 may bind to capsid vertices [Zhou, Z. H., Chen, D. H., Jakana, J., Rixon, F. J. & Chiu, W. (1999) J. Virol. 73, 3210-3218; Klupp, B. G., Fuchs, W., Granzow, H., Nixdorf, R. & Mettenleiter, T. C. (2002) J. Virol. 76, 3065-3071].