Intracellular localization of Equine herpesvirus type 1 tegument protein VP22

•We examined intracellular localization of EHV-1 VP22 (EVP22) in infected cells and transfected cells.•EVP22 localizes cytoplasm and nucleus independently of other viral proteins.•The region 154–188 aa of EVP22 is required for nuclear translocation.

Intracellular localization of Equine herpesvirus type 1 (EHV-1) tegument protein VP22 was examined by using a plasmid that expressed VP22 fused with an enhanced green fluorescent protein (EGFP). Also a recombinant EHV-1 expressing VP22 fused with a red fluorescent protein (mCherry) was constructed to observe the localization of VP22 in infected cells. When EGFP-fused VP22 was overexpressed in the cells, VP22 localized in the cytoplasm and nucleus. Live cell imaging suggested that the fluorescently tagged VP22 also localized in the cytoplasm and nucleus. These results show that VP22 localizes in the cytoplasm and nucleus independently of other viral proteins. Experiments with truncation mutants of pEGFP-VP22 suggested that 154–188 aa might be the nuclear localization signal of EHV-1 VP22.