Translocation of incoming pseudorabies virus capsids to the cell nucleus is delayed in the absence of tegument protein pUL37.
After fusion of the envelope of herpesvirus particles with the host cell plasma membrane, incoming nucleocapsids are transported to nuclear pores. Inner tegument proteins pUL36, pUL37, and pUS3 remain attached to the nucleocapsid after entry and therefore might mediate interactions between the nucleocapsid and cellular microtubule-associated motor proteins during transport. To assay for the role of pUL37 in this process, we constructed a pUL37-deleted pseudorabies virus mutant, PrV-DeltaUL37/UL35GFP, which expresses a fusion protein of green fluorescent protein (GFP) and the nonessential small capsid protein pUL35, resulting in the formation of fluorescently labeled capsids. Confocal laser-scanning microscopy of rabbit kidney cells infected with PrV-DeltaUL37/UL35GFP revealed that, whereas penetration was not affected in the absence of pUL37, nuclear translocation of incoming particles was delayed by approximately 1 h compared to PrV-UL35GFP, but not abolished. In contrast, phenotypically complemented pUL37-containing virions of PrV-DeltaUL37/UL35GFP exhibited wild type-like entry kinetics. Thus, the presence of pUL37 is required for rapid nuclear translocation of incoming nucleocapsids.
Krautwald
Mirjam
M
Institute of Molecular Biology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald-Insel Riems, Germany.
Fuchs
Walter
W
Klupp
Barbara G
BG
Mettenleiter
Thomas C
TC
D000818:Animals; D002213:Capsid / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000821:virology*; D017353:Gene Deletion; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000502:physiology*; D018613:Microscopy, Confocal; D021381:Protein Transport; D011817:Rabbits; D013194:Staining and Labeling / Q000379:methods; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism*
D015678:Viral Structural Proteins; D049452:Green Fluorescent Proteins