title: Probing of the nuclear import and export signals and subcellular transport mechanism of varicella-zoster virus tegument protein open reading frame 10.
abstract: Varicella-zoster virus open reading frame 10 (ORF10), a tegument protein present in the virion, is a member of the alphaherpesvirus UL48 gene family that shares considerable amino acid sequence homology with the UL48 prototype, herpes simplex virus type 1 VP16. VP16 serves multiple functions, including transcriptional activation of viral immediate-early genes. Furthermore, VP16 has been shown to be involved in some aspects of virus assembly and/or maturation. However, little is known concerning the function of ORF10. Here, we found that transient expression of ORF10 fused to enhanced yellow fluorescent protein (EYFP) in COS-7 cells showed the predominantly nuclear localization in the absence of other viral proteins. By constructing a series of ORF10 variants fused to EYFP, a bona fide bipartite nuclear localization signal of ORF10 was, for the first time, determined and mapped to amino acids (aa) 302-347. Additionally, the nuclear export signal (NES) was identified and found to be in a leucine-rich region at aa 226-236. Finally, ORF10 was demonstrated to be targeted to the cytoplasm through the functional NES by chromosomal region maintenance 1-dependent pathway, and to the nucleus via Ran and importin β1-dependent pathway that does not require importin α5.
authors.affiliation: State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuchang, Wuhan, China.
authors.forename: Mingsheng
authors.initials: M
authors.lastname: Cai
authors.forename: Shuai
authors.initials: S
authors.lastname: Wang
authors.forename: Jing
authors.initials: J
authors.lastname: Long
authors.forename: Chunfu
authors.initials: C
authors.lastname: Zheng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D019556:COS Cells; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D002522:Chlorocebus aethiops; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D028884:Karyopherins / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D049790:Nuclear Export Signals / Q000235:genetics*; D019913:Nuclear Localization Signals / Q000737:chemistry / Q000235:genetics*; D021381:Protein Transport; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism*; D028961:beta Karyopherins / Q000235:genetics / Q000378:metabolism; D020931:ran GTP-Binding Protein / Q000235:genetics / Q000378:metabolism; D000097606:Exportin 1 Protein
chemical_list: D001426:Bacterial Proteins; D028884:Karyopherins; D008164:Luminescent Proteins; D049790:Nuclear Export Signals; D019913:Nuclear Localization Signals; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D015678:Viral Structural Proteins; D028961:beta Karyopherins; D020931:ran GTP-Binding Protein; D000097606:Exportin 1 Protein; C090372:ORF 10 protein, varicella-zoster virus; C054562:yellow fluorescent protein, Bacteria