Cellular protein binds to sequences near the 5' terminus of potato virus X RNA that are important for virus replication.
Cellular protein binds to sequences near the 5' terminus of potato virus X RNA that are important for virus replication.
复制标题
细胞蛋白与马铃薯病毒 X RNA 5 末端附近的序列结合，这对病毒复制很重要。
DOI：
10.1006/viro.2002.1559
复制
发表时间：
2002
期刊：
影响因子：
3.7
通讯作者：
Hemenway,Cynthia
中科院分区：
文献类型：
--
作者：
Kim,Kook-Hyung;Kwon,Sun-Jung;Hemenway,Cynthia
马铃薯X病毒(PVX)基因组RNA的5‘-非翻译区(NTR)序列含有基因组和亚基因组RNA积累所需的几个调控元件。为了研究细胞蛋白是否与这些元件结合，我们用来自PVX 5‘-NTR的原生质体蛋白质提取物和RNA序列进行了凝胶迁移率改变分析(EMSA)。这些分析表明，PVX正链RNA的5‘端与细胞蛋白形成了络合物。对不同缺失的PVX RNA形成的复合体的紫外光交联研究表明，一个54 kDa的细胞蛋白(P54)与PVX RNA 5‘端的1-46位核苷酸结合。在该46-nT区域引入的定点突变进一步表明，位于nt10-13的ACCA序列元件对于最佳结合是重要的。此外，降低模板RNA对细胞因子亲和力的突变减少了PVX正链RNA在原生质体中的积累。这些研究表明，p54可能通过与病毒基因组RNA的5‘末端结合而在PVX RNA复制中发挥作用。
The sequences in the 5′-nontranslated region (NTR) of Potato virus X (PVX) genomic RNA were previously reported to contain several regulatory elements that are required for genomic and subgenomic RNA accumulation. To investigate whether cellular proteins bind to these elements, we conducted electrophoretic mobility shift assays (EMSA) with protoplast protein extracts and RNA sequences from the PVX 5′-NTR. These analyses showed that the 5′ region of PVX positive-strand RNA formed complexes with cellular proteins. UV cross-linking studies of complexes formed with various deletions of the PVX RNA indicated that a 54-kDa cellular protein (p54) was bound to nt 1–46 at the 5′ terminus of PVX RNA. Site-directed mutations introduced within this 46-nt region further indicated that an ACCA sequence element located at nt 10–13 was important for optimal binding. In addition, mutations that decreased the affinity of the template RNA for the cellular factor decreased PVX plus-strand RNA accumulation in protoplasts. These studies suggest that the p54 may function in PVX RNA replication by binding to the 5′ terminus of the viral genomic RNA.