Nuclear import of prototype foamy virus transactivator Bel1 is mediated by KPNA1, KPNA6 and KPNA7.
Bel1, a transactivator of the prototype foamy virus (PFV), plays pivotal roles in the replication of PFV. Previous studies have demonstrated that Bel1 bears a nuclear localization signal (NLS); however, its amino acid sequence remains unclear and the corresponding adaptor importins have not yet been identified. In this study, we inserted various fragments of Bel1 into an EGFP-GST fusion protein and investigated their subcellular localization by fluorescence microscopy. We found that the 215PRQKRPR221 fragment, which accords with the consensus sequence K(K/R)X(K/R) of the monopartite NLS, directed the nuclear translocation of Bel1. Point mutation experiments revealed that K218, R219 and R221 were essential for the nuclear localization of Bel1. The results of GST pull-down assay revealed that the Bel1 peptide 215-221, which bears the NLS, interacted with the nucleocytoplasmic transport receptors, karyopherin alpha 1 (importin alpha 5) (KPNA1), karyopherin alpha 6 (importin alpha 7) (KPNA6) and karyopherin alpha 7 (importin alpha 8) (KPNA7). Finally, in vitro nuclear import assays demonstrated that KPNA1, KPNA6 or KPNA7, along with other necessary nuclear factors, caused Bel1 to localize to the nucleus. Thus, the findings of our study indicate that KPNA1, KPNA6 and KPNA7 are involved in Bel1 nuclear distribution.
Clinical Laboratory, Tianjin First Center Hospital, Tianjin 300192, P.R. China.
Jihui
J
Duan
Clinical Laboratory, Tianjin First Center Hospital, Tianjin 300192, P.R. China.
Zhiqin
Z
Tang
Clinical Laboratory, Tianjin First Center Hospital, Tianjin 300192, P.R. China.
Hong
H
Mu
Clinical Laboratory, Tianjin First Center Hospital, Tianjin 300192, P.R. China.
Guojun
G
Zhang
D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D002463:Cell Membrane Permeability; D002467:Cell Nucleus / Q000378:metabolism*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D028884:Karyopherins / Q000378:metabolism*; D019913:Nuclear Localization Signals / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D012191:Retroviridae Proteins / Q000737:chemistry / Q000378:metabolism*; D016092:Spumavirus / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism*
D028884:Karyopherins; D019913:Nuclear Localization Signals; D012191:Retroviridae Proteins; D015534:Trans-Activators; C074061:bel1 protein, Human foamy virus