Fusion of enhanced green fluorescent protein to the pseudorabies virus axonal sorting protein Us9 blocks anterograde spread of infection in mammalian neurons.
Pseudorabies virus encodes a membrane protein (Us9) that is essential for the axonal sorting of virus particles within neurons and anterograde spread in the mammalian nervous system. Enhanced green fluorescent protein (GFP)-tagged Us9 mimicked the trafficking properties of the wild-type protein in nonneuronal cells. We constructed a pseudorabies virus strain that expressed Us9-GFP and tested its spread capabilities in the rat visual system and in primary neuronal cultures. We report that Us9-EGFP does not promote anterograde spread of infection and may disrupt packing of viral membrane proteins in lipid rafts, an essential step for Us9-mediated axonal sorting.
Lyman
M G
MG
314 Schultz Laboratory, Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA.
Curanovic
D
D
Brideau
A D
AD
Enquist
L W
LW
D000818:Animals; D001369:Axons / Q000378:metabolism*; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism*; D008297:Male; D021962:Membrane Microdomains / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000821:virology; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D011557:Pseudorabies / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D047908:Intracellular Signaling Peptides and Proteins; D008074:Lipoproteins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; C491630:US9 protein, Suid herpesvirus 1; D014764:Viral Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins