Tyc K, Steitz J A
Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT 06510.
EMBO J. 1989 Oct;8(10):3113-9. doi: 10.1002/j.1460-2075.1989.tb08463.x.
Using anti-(U3)RNP autoantibodies, we have isolated and characterized two additional small nucleolar RNAs from HeLa cells, which are less abundant than U3 RNA. Both RNAs possess a trimethylguanosine cap as judged by precipitation with anti-TMG antibody, but are not precipitated by either anti-Sm or anti-La antibodies. In addition, both RNAs are not precipitable by anti-Th serum, which recognizes another nucleolar RNP autoantigen. Sequence analysis revealed that one of these RNAs, 136 nucleotides long, is the human U8 homolog; while the other, 105 nucleotides long, represents a novel species which we designate U13. Both RNAs share with U3 two conserved sequences (boxes C and D). The role of one or both of these boxes in binding the common 34 kd antigenic protein, otherwise known as fibrillarin, is discussed. Fractionation of HeLa cells revealed that U8 and U13, like U3, reside in the nucleolus. In glycerol gradients both RNAs cosediment with larger structures possibly representing ribosomal precursors. We propose that U3, U8 and U13 comprise a new subset of mammalian snRNPs whose roles in ribosome biogenesis are discussed.
利用抗(U3)RNP自身抗体,我们从HeLa细胞中分离并鉴定出另外两种小分子核仁RNA,它们的丰度低于U3 RNA。通过抗三甲基鸟苷(TMG)抗体沉淀判断,这两种RNA都具有三甲基鸟苷帽,但不能被抗Sm或抗La抗体沉淀。此外,这两种RNA也不能被识别另一种核仁RNP自身抗原的抗Th血清沉淀。序列分析表明,其中一种RNA长136个核苷酸,是人类U8的同源物;而另一种长105个核苷酸,代表一个新的种类,我们将其命名为U13。这两种RNA与U3都有两个保守序列(C盒和D盒)。讨论了这些盒子中的一个或两个在结合常见的34 kd抗原蛋白(即原纤维蛋白)中的作用。HeLa细胞分级分离显示,U8和U13与U3一样,存在于核仁中。在甘油梯度中,这两种RNA都与可能代表核糖体前体的较大结构共沉降。我们提出,U3、U8和U13构成了哺乳动物小核核糖核蛋白(snRNP)的一个新亚组,并讨论了它们在核糖体生物发生中的作用。