Lee S I, Steitz J A
Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, Connecticut 06510.
J Virol. 1990 Aug;64(8):3905-15. doi: 10.1128/JVI.64.8.3905-3915.1990.
Marmoset T lymphocytes transformed by herpesvirus saimiri contain a set of five virally encoded U RNAs called HSUR1 through HSUR5. HSUR genes have been individually transfected into a nonlymphoid, nonsimian cell line (HeLa cells) in the absence of any other coding regions of the herpesvirus saimiri genome. The levels of HSUR1 through HSUR4 in HeLa transient-expression systems are comparable to those found in virally transformed T cells (23 to 91%). In contrast, HSUR5 is expressed at ninefold-higher levels in transfected HeLa cells. Immunoprecipitation experiments show that HSURs expressed in transfected cells bind proteins with Sm determinants and acquire a 5' trimethylguanosine cap structure, as they do in transformed T cells. HSUR1 or HSUR4 particles from transfected HeLa cells migrate between 10S and 15S in velocity gradients, identical to the sedimentation of "monoparticles" produced in virally transformed lymphocytes. We conclude from these transfection experiments that no other herpesvirus saimiri or host-cell-specific gene products appear to be required for efficient expression of the HSUR genes or for subsequent assembly of the viral U RNAs into small nuclear ribonucleoprotein particles. In lymphocytes transformed by herpesvirus saimiri, HSUR small nuclear ribonucleoprotein particles are involved in higher-order complexes that sediment between 20S and 25S. HSUR1, HSUR2, and HSUR5 dissociate from such complexes upon incubation at 30 degrees C, whereas the complex containing HSUR4 is stable to incubation.
被猴疱疹病毒转化的狨猴T淋巴细胞含有一组五个病毒编码的U RNA,称为HSUR1至HSUR5。在没有猴疱疹病毒基因组的任何其他编码区的情况下,HSUR基因已被单独转染到非淋巴细胞、非猴细胞系(HeLa细胞)中。在HeLa瞬时表达系统中,HSUR1至HSUR4的水平与在病毒转化的T细胞中发现的水平相当(23%至91%)。相比之下,HSUR5在转染的HeLa细胞中的表达水平高九倍。免疫沉淀实验表明,转染细胞中表达的HSUR与具有Sm决定簇的蛋白质结合,并获得5'三甲基鸟苷帽结构,就像它们在转化的T细胞中一样。来自转染HeLa细胞的HSUR1或HSUR4颗粒在速度梯度中迁移在10S和15S之间,与病毒转化的淋巴细胞中产生的“单颗粒”的沉降相同。我们从这些转染实验得出结论,高效表达HSUR基因或随后将病毒U RNA组装成小核核糖核蛋白颗粒似乎不需要其他猴疱疹病毒或宿主细胞特异性基因产物。在被猴疱疹病毒转化的淋巴细胞中,HSUR小核核糖核蛋白颗粒参与沉降在20S和25S之间的高阶复合物。在30℃孵育时,HSUR1、HSUR2和HSUR5从这种复合物中解离,而含有HSUR4的复合物在孵育时是稳定的。