Characterization of the nuclear and nucleolar localization signals of bovine herpesvirus-1 infected cell protein 27.
Bovine herpesvirus-1 infected cell protein 27 (BICP27) was detected predominantly in the nucleolus. The open reading frame of BICP27 was fused with the enhanced yellow fluorescent protein (EYFP) gene to investigate its subcellular localization in live cells and BICP27 was able to direct monomeric, dimeric or trimeric EYFP exclusively to the nucleolus. By constructing a series of deletion mutants, the putative nuclear localization signal (NLS) and nucleolar localization signal (NoLS) were mapped to (81)RRAR(84) and (86)RPRRPRRRPRRR(97) respectively. Specific deletion of the putative NLS, NoLS or both abrogated nuclear localization, nucleolar localization or both respectively. Furthermore, NLS was able to direct trimeric EYFP predominantly to the nucleus but excluded from the nucleolus, whereas NoLS targeted trimeric EYFP primarily to the nucleus, and enriched in the nucleolus with faint staining in the cytoplasm. NLS+NoLS directed trimeric EYFP predominantly to the nucleolus with faint staining in the nucleus. Moreover, deletion of NLS+NoLS abolished the transactivating activity of BICP27 on gC promoter, whereas deletion of either NLS or NoLS did not. The study demonstrated that BICP27 is a nucleolar protein, adding BICP27 to the growing list of transactivators which localize to the nucleolus.
Guo
Hong
H
State Key Laboratory of Virology, Molecular Virology and Viral Immunology Research Group, Wuhan Institute of Virology, Chinese Academy of Sciences, 44 Xiaohongshan, Wuchang, Wuhan, Hubei 430071, PR China.
Ding
Qiong
Q
Lin
Fusen
F
Pan
Weiwei
W
Lin
Jianyin
J
Zheng
Alan C
AC
D000818:Animals; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D002466:Cell Nucleolus / Q000737:chemistry*; D017930:Genes, Reporter; D000882:Haplorhini; D007242:Herpesvirus 1, Bovine / Q000254:growth & development*; D017874:Immediate-Early Proteins / Q000235:genetics* / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism*; D021382:Protein Sorting Signals; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion
C099232:BICP27 protein, Bovine herpesvirus 1; D001426:Bacterial Proteins; D017874:Immediate-Early Proteins; D008164:Luminescent Proteins; D010750:Phosphoproteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria
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