Nunes-Correia Isabel, Rodríguez Javier M, Eulálio Ana, Carvalho Ana L, Citovsky Vitaly, Simões Sérgio, Faro Carlos, Salas María L, Pedroso de Lima Maria C
Centro de Neurociências e Biologia Celular, Universidade de Coimbra, 3004-517 Coimbra, Portugal.
Vet Microbiol. 2008 Jul 27;130(1-2):47-59. doi: 10.1016/j.vetmic.2007.12.010. Epub 2007 Dec 28.
African swine fever virus (ASFV), a large enveloped DNA-containing virus, infects domestic and wild pigs, and multiplies in soft ticks, causing an economically relevant hemorrhagic disease. Evaluation of the nuclear import ability of ASFV p10 protein was the major purpose of the present work. Two approaches were used to determine if p10 protein is imported into the nucleus by an active process: a yeast-based nuclear import assay and the determination of the subcellular localization of p10 protein in mammalian cells by fluorescence microscopy. The results obtained clearly demonstrate that p10 protein is actively imported into the nucleus, both in yeast and mammalian cells. Experiments aiming at identifying the critical residues responsible for the nuclear import of ASFV p10 protein indicate that the amino acids comprised between the positions 71 and 77 are important, although not sufficient, for the protein active nuclear import. In ASFV-infected cells, the p10 protein strongly accumulates in the nucleus at late times post-infection, indicating that p10 protein may accomplish an important function inside the nucleus during the late phase of the viral replication cycle.
非洲猪瘟病毒(ASFV)是一种大型的含包膜DNA病毒,可感染家猪和野猪,并在软蜱中繁殖,引发一种具有经济影响的出血性疾病。评估ASFV p10蛋白的核输入能力是本研究的主要目的。采用了两种方法来确定p10蛋白是否通过主动过程被导入细胞核:基于酵母的核输入测定法以及通过荧光显微镜确定p10蛋白在哺乳动物细胞中的亚细胞定位。所获得的结果清楚地表明,p10蛋白在酵母和哺乳动物细胞中均能被主动导入细胞核。旨在鉴定负责ASFV p10蛋白核输入的关键残基的实验表明,第71至77位之间的氨基酸对于该蛋白的主动核输入很重要,尽管并不充分。在ASFV感染的细胞中,p10蛋白在感染后期强烈积聚在细胞核中,这表明p10蛋白可能在病毒复制周期的后期在细胞核内发挥重要功能。