Lee Hye-Ra, Ahn Jin-Hyun
School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea.
Department of Molecular Cell Biology, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon, Kyonggido 440-746, Korea.
J Gen Virol. 2004 Aug;85(Pt 8):2149-2154. doi: 10.1099/vir.0.79954-0.
Sumoylation of the major immediate-early IE2 protein of human cytomegalovirus has been shown to increase transactivation activity in target reporter gene assays. This study examined the role of IE2 sumoylation in viral infection. A Towne strain-based bacterial artificial chromosome clone was generated encoding a mutated form of the IE2 protein with Lys-->Arg substitutions at positions 175 and 180, the two major sumoylation sites. When human fibroblast (HF) cells were infected with the reconstituted mutant virus, (i) viral growth kinetics, (ii) the accumulation of IE1 (UL123), IE2 (UL122), p52 (UL44) and pp65 (UL83) proteins and (iii) the relocalization of the cellular small ubiquitin-like modifier (SUMO)-1, p53 and proliferating cell nuclear antigen proteins into viral DNA replication compartments were comparable with those of the wild-type and the revertant virus. The data demonstrate that sumoylation of IE2 is not essential for virus growth in cultured HF cells.
人巨细胞病毒主要立即早期IE2蛋白的SUMO化已被证明在靶报告基因检测中可增加反式激活活性。本研究检测了IE2 SUMO化在病毒感染中的作用。构建了一个基于Towne株的细菌人工染色体克隆,其编码IE2蛋白的突变形式,在两个主要SUMO化位点175和180位的赖氨酸被替换为精氨酸。当用人成纤维细胞(HF)感染重组突变病毒时,(i)病毒生长动力学、(ii)IE1(UL123)、IE2(UL122)、p52(UL44)和pp65(UL83)蛋白的积累以及(iii)细胞小泛素样修饰物(SUMO)-1、p53和增殖细胞核抗原蛋白重新定位到病毒DNA复制区室的情况与野生型和回复病毒相当。数据表明,IE2的SUMO化对于培养的HF细胞中的病毒生长不是必需的。