Glickman J N, Howe J G, Steitz J A
Howard Hughes Medical Institute, Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06510-8024.
J Virol. 1988 Mar;62(3):902-11. doi: 10.1128/JVI.62.3.902-911.1988.
The ribonucleoprotein (RNP) particles containing the Epstein-Barr virus-associated small RNAs EBER1 and EBER2 were analyzed to determine their RNA secondary structures and sites of RNA-protein interaction. The secondary structures were probed with nucleases and by chemical modification with single-strand-specific reagents, and the sites of modification or cleavage were mapped by primer extension. These data were used to develop secondary structures for the two RNAs, and likely sites of close RNA-protein contact were identified by comparing modification patterns for naked RNA and RNA in RNP particles. In addition, sites of interaction between each Epstein-Barr virus-encoded RNA (EBER) and the La antigen were identified by analyzing RNA fragments resistant to digestion by RNase A or T1 after immunoprecipitation by an anti-La serum sample from a lupus patient. Our results confirm earlier findings that the La protein binds to the 3' terminus of each molecule. Possible functions for the EBER RNPs are discussed.
对含有爱泼斯坦-巴尔病毒相关小RNA EBER1和EBER2的核糖核蛋白（RNP）颗粒进行分析，以确定其RNA二级结构和RNA-蛋白质相互作用位点。用核酸酶和单链特异性试剂进行化学修饰来探测二级结构，通过引物延伸对修饰或切割位点进行定位。这些数据用于构建这两种RNA的二级结构，并通过比较裸RNA和RNP颗粒中RNA的修饰模式来确定可能的紧密RNA-蛋白质接触位点。此外，通过分析来自狼疮患者的抗La血清样本免疫沉淀后对RNase A或T1消化有抗性的RNA片段，确定了每个爱泼斯坦-巴尔病毒编码RNA（EBER）与La抗原之间的相互作用位点。我们的结果证实了早期的发现，即La蛋白与每个分子的3'末端结合。文中还讨论了EBER RNP的可能功能。
