Krauer Kenia G, Buck Marion, Belzer Deanna K, Flanagan James, Chojnowski Grace M, Sculley Tom B
Queensland Institute of Medical Research and ACITHN University of Queensland, Brisbane 4029, Queensland, Australia.
Virology. 2004 Jan 5;318(1):280-94. doi: 10.1016/j.virol.2003.09.032.
The Epstein-Barr virus nuclear antigen (EBNA)-6 protein is essential for Epstein-Barr virus (EBV)-induced immortalization of primary human B-lymphocytes in vitro. In this study, fusion proteins of EBNA-6 with green fluorescent protein (GFP) have been used to characterize its nuclear localization and organization within the nucleus. EBNA-6 associates with nuclear structures and in immunofluorescence demonstrate a punctate staining pattern. Herein, we show that the association of EBNA-6 with these nuclear structures was maintained throughout the cell cycle and with the use of GFP-E6 deletion mutants, that the region amino acids 733-808 of EBNA-6 contains a domain that can influence the association of EBNA-6 with these nuclear structures. Co-immunofluorescence and confocal analyses demonstrated that EBNA-6 and EBNA-3 co-localize in the nucleus of cells. Expression of EBNA-6, but not EBNA-3, caused a redistribution of nuclear survival of motor neurons protein (SMN) to the EBNA-6 containing nuclear structures resulting in co-localization of SMN with EBNA-6.
爱泼斯坦-巴尔病毒核抗原(EBNA)-6蛋白对于爱泼斯坦-巴尔病毒(EBV)在体外诱导原代人B淋巴细胞永生化至关重要。在本研究中,EBNA-6与绿色荧光蛋白(GFP)的融合蛋白已被用于表征其在细胞核内的定位和组织。EBNA-6与核结构相关联,在免疫荧光中呈现点状染色模式。在此,我们表明EBNA-6与这些核结构的关联在整个细胞周期中都得以维持,并且通过使用GFP-E6缺失突变体发现,EBNA-6的733-808位氨基酸区域包含一个可影响EBNA-6与这些核结构关联的结构域。共免疫荧光和共聚焦分析表明,EBNA-6和EBNA-3在细胞核中共定位。EBNA-6而非EBNA-3的表达导致运动神经元存活蛋白(SMN)在细胞核内重新分布至含有EBNA-6的核结构,从而导致SMN与EBNA-6共定位。