N-terminal of Lprotein of vesicular stomatitis virus contains a new signal sequence  

作　　者：NIE Yuchun, KE Yeyan, WANG Zai, YU Xiang, DENG Hongkui & DING Mingxiao Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871, China Correspondence should be addressed to Ding Mingxiao (e-mail: [e-mail]) 

出　　处：《Chinese Science Bulletin》2003年第13期1352-1357,共6页

基　　金：supported by the Major State Basic Research Program of China(Grant No.G1999011900);an Outstanding Young Investigator from the National Natural Science Foundation of China(Grant No.30125022).

摘　　要：The L protein (241 kD) of vesicular stomatitis virus (VSV) is the most important subunit of the replication complex. The existence of specific localization signal in the L protein was investigated by making recombinant constructs expressing truncated mutants of the L protein fused to green fluorescent protein (GFP) in transient transfection assays. The chimeric genes encoding varied N-terminal of L and GFP gene were put under the control of T7 promoter or CMV promoter. The fusion proteins were transiently expressed in BHK-21, COS-7, CHO or Hep G2 cells. When more than 120 residues were deleted or only 96 residues were kept on the N-terminal, the fusion proteins were shown to be distributed throughout the cells, cytoplasm and nucleus under the confocal microscope. However, other chimeric proteins with 120 or more amino acids were dotted and distributed in the perinuclear regions. And the fusion protein with 96120 aa has the similar distribution. A thirteen-residue peptide QGYSFLHEVDKEA (108120) was identified as localization signal, whose function would be absolutely distributed with the deficiency of D or V. Our results show that there is an independent localizing signal in N-terminal domain of L protein of VSV and this functional signal is conserved in different cell lines.The L protein (241 kD) of vesicular stomatitis virus (VSV) is the most important subunit of the replication complex. The existence of specific localization signal in the L protein was investigated by making recombinant constructs expressing truncated mutants of the L protein fused to green fluorescent protein (GFP) in transient transfection assays. The chimeric genes encoding varied N-terminal of L and GFP gene were put under the control of T7 promoter or CMV promoter. The fusion proteins were transiently expressed in BHK-21, COS-7, CHO or Hep G2 cells. When more than 120 residues were deleted or only 96 residues were kept on the N-terminal, the fusion proteins were shown to be distributed throughout the cells, cytoplasm and nucleus under the confocal microscope. However, other chimeric proteins with 120 or more amino acids were dotted and distributed in the perinuclear regions. And the fusion protein with 96120 aa has the similar distribution. A thirteen-residue peptide QGYSFLHEVDKEA (108120) was identified as localization signal, whose function would be absolutely distributed with the deficiency of D or V. Our results show that there is an independent localizing signal in N-terminal domain of L protein of VSV and this functional signal is conserved in different cell lines.

关 键 词：L蛋白质 水泡性口膜炎病毒 VSV 基因编码 绿色荧光蛋白质 

分 类 号：R373[医药卫生—病原生物学]