García-Montalvo Beatriz María, Medina Fernando, del Angel Rosa M
Departamento de Patología Experimental, Centro de Investigación y de Estudios Avanzados del IPN, Av. I.P.N. 2508, Col. San Pedro Zacatenco, México, D.F. C.P. 07360, Mexico.
Virus Res. 2004 Jun 15;102(2):141-50. doi: 10.1016/j.virusres.2004.01.024.
In vitro replication of dengue virus requires the presence of cis-acting elements within the 5' end and the 3' UTR of the viral genome. Some, like the putative cyclization sites (PCS), may promote interaction at both ends of the viral genome. To investigate whether viral or cellular proteins could be involved in this interaction, UV-induced cross-linking assays using extracts from the monocytic cell line U937 were performed. Our data demonstrate that the 5' end and the 3' UTR with the PCS interact with five cellular proteins with the same molecular weight. When both regions were differentially labeled, with biotin and 32P, respectively, the interaction of at least seven proteins with both ends could be demonstrated. Immunoprecipitation assays also demonstrate that La protein binds to the 5' end and with the 3' UTR. Moreover, these proteins also interact with the nonstructural proteins NS5 and NS3. The role of the NS5-La and NS3-La interaction in U937 cells remains to be established.
登革病毒的体外复制需要病毒基因组5'端和3'非翻译区（UTR）内存在顺式作用元件。其中一些元件，如假定的环化位点（PCS），可能促进病毒基因组两端的相互作用。为了研究病毒蛋白或细胞蛋白是否参与这种相互作用，我们使用单核细胞系U937的提取物进行了紫外线诱导的交联试验。我们的数据表明，带有PCS的5'端和3'UTR与五种分子量相同的细胞蛋白相互作用。当两个区域分别用生物素和32P进行差异标记时，可以证明至少有七种蛋白与两端相互作用。免疫沉淀试验也表明，La蛋白与5'端和3'UTR结合。此外，这些蛋白还与非结构蛋白NS5和NS3相互作用。NS5-La和NS3-La相互作用在U937细胞中的作用仍有待确定。
