TY - JOUR
T1 - Molecular anatomy of subcellular localization of HSV-1 tegument protein US11 in living cells
AU - Xing, Junji
AU - Wu, Fuqing
AU - Pan, Weiwei
AU - Zheng, Chunfu
N1 - Funding Information:
This work was supported by grants from the Major State Basic Research Development Program of China (973 Program) (2010CB530105); the Start-up Fund of the Hundred Talents Program of the Chinese Academy of Sciences (20071010-141); the National Natural Science Foundation of China (30870120) and Hubei Province Natural Science Foundation of Innovation Groups Project (2008CDA013). We thank Dr. Yasushi Kawaguchi for the generous gift pYEbac102, Dr. Yoshinari Yasuda for the generous gift plasmid pGEX-6p-1-Q69L Ran, Dr. Chris Basler for the generous gift plasmid DN-importin α5, Dr. Haitao Guo for the generous gift plasmid DN-importin β and Dr. Gillian Elliott for the generous gift plasmid pRev-NES-EGFP.
PY - 2010/10
Y1 - 2010/10
N2 - The herpes simplex virus type I (HSV-1) US11 protein is an RNA-binding multifunctional regulator that specifically and stably associates with nucleoli. Although the C-terminal part of US11 was responsible for its nucleolar localization, the precise nucleolar localization signal (NoLS) and nuclear export signal (NES) of US11 and its nuclear import and export mechanisms are still elusive. In this study, fluorescence microscopy was employed to investigate the subcellular localization of US11 and characterize its transport mechanism in living cells. By constructing a series of deletion mutants fused with enhanced yellow fluorescent protein (EYFP), three novel NoLSs of US11 were for the first time mapped to amino acids 84-125, 126-152, and 89-146, respectively. Additionally, the NES was identified to locate between amino acids 89 and 119. Furthermore, the US11 protein was demonstrated to target to the cytoplasm through the NES by chromosomal region maintenance 1 (CRM1)-independent pathway, and to the nucleolus through Ran and importin β-dependent mechanism that does not require importin α5.
AB - The herpes simplex virus type I (HSV-1) US11 protein is an RNA-binding multifunctional regulator that specifically and stably associates with nucleoli. Although the C-terminal part of US11 was responsible for its nucleolar localization, the precise nucleolar localization signal (NoLS) and nuclear export signal (NES) of US11 and its nuclear import and export mechanisms are still elusive. In this study, fluorescence microscopy was employed to investigate the subcellular localization of US11 and characterize its transport mechanism in living cells. By constructing a series of deletion mutants fused with enhanced yellow fluorescent protein (EYFP), three novel NoLSs of US11 were for the first time mapped to amino acids 84-125, 126-152, and 89-146, respectively. Additionally, the NES was identified to locate between amino acids 89 and 119. Furthermore, the US11 protein was demonstrated to target to the cytoplasm through the NES by chromosomal region maintenance 1 (CRM1)-independent pathway, and to the nucleolus through Ran and importin β-dependent mechanism that does not require importin α5.
KW - Herpes simplex virus 1 US11 protein
KW - Importin
KW - Nuclear export signal
KW - Nucleolar localization signal
KW - Ran-GTP
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U2 - 10.1016/j.virusres.2010.07.009
DO - 10.1016/j.virusres.2010.07.009
M3 - Article
C2 - 20633584
AN - SCOPUS:77956231403
SN - 0168-1702
VL - 153
SP - 71
EP - 81
JO - Virus Research
JF - Virus Research
IS - 1
ER -