Identification of molecular determinants for the nuclear import of pseudorabies virus UL31

• PRV UL31 was confirmed to exclusively target to the nucleus.
• The nuclear import of PRV UL31 was found to be dependent on the Ran-, importin α1-, α3-, α5-, α7-, α1- and transportin-1-mediated pathway.
• These results would open up new avenues for depicting the biological functions of UL31 during PRV infection.

Herpes simplex virus 1 (HSV-1) UL31 is a multifunctional protein and important for HSV-1 infection. Pseudorabies virus (PRV) UL31 is a late protein homologous to HSV-1 UL31. Previous studies showed that PRV UL31 is predominantly localized to nucleus, however, the molecular determinants for its nuclear import were unclear to date. Here, by utilizing live cells fluorescent microscopy, UL31 fused with enhanced yellow fluorescent protein was transiently expressed in live cells and confirmed to exclusively target to the nucleus in the absence of other viral proteins. Furthermore, the nuclear import of UL31 was found to be dependent on the Ran-, importin α1-, α3-, α5-, α7-, β1-and transportin-1-mediated pathway. Therefore, these results would open up new avenues for depicting the biological functions of UL31 during PRV infection.