Tycowski K T, Shu M D, Steitz J A
Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, Connecticut 06536-0812.
Genes Dev. 1993 Jul;7(7A):1176-90. doi: 10.1101/gad.7.7a.1176.
A human small nucleolar RNA, identified previously in HeLa cells by anti-fibrillarin autoantibody precipitation and termed RNA X, has been characterized. It comprises two uridine-rich variants (148 and 146 nucleotides), which we refer to as snRNA U15A and U15B. Secondary structure models predict for both variants a U15-specific stem-loop structure, as well as a new structural motif that contains conserved sequences and can also be recognized in the other fibrillarin-associated nucleolar snRNAs, U3, U14, and RNA Y. The single-copy gene for human U15A has been found unexpectedly to reside in intron 1 of the ribosomal protein S3 gene; the U15A sequence appears on the same strand as the S3 mRNA and does not exhibit canonical transcription signals for nuclear RNA polymerases. U15A RNA is processed in vitro from S3 intron 1 transcripts to yield the correct 5' end with a 5'-monophosphate; the in vitro system requires ATP for 3' cleavage, which occurs a few nucleotides downstream of the mature end. The production of a single primary transcript specifying the mRNA for a ribosomal or nucleolar protein and a nucleolar snRNA may constitute a general mechanism for balancing the levels of nucleolar components in vertebrate cells.
一种人类小核仁RNA已被鉴定,它先前在HeLa细胞中通过抗原纤维蛋白自身抗体沉淀法被发现,并被命名为RNA X。它由两个富含尿苷的变体(148和146个核苷酸)组成,我们将其称为小核仁RNA U15A和U15B。二级结构模型预测这两个变体都有一个U15特异性的茎环结构,以及一个包含保守序列的新结构基序,该基序也能在其他与原纤维蛋白相关的核仁小核仁RNA、U3、U14和RNA Y中被识别。人类U15A的单拷贝基因意外地位于核糖体蛋白S3基因的内含子1中;U15A序列与S3 mRNA位于同一条链上,并且没有显示出核RNA聚合酶的典型转录信号。U15A RNA在体外从S3内含子1转录本加工而成,以产生具有5'-单磷酸的正确5'末端;体外系统需要ATP进行3'切割,切割发生在成熟末端下游几个核苷酸处。产生一个单一的初级转录本,该转录本指定核糖体或核仁蛋白的mRNA以及核仁小核仁RNA,可能构成一种平衡脊椎动物细胞核仁成分水平的普遍机制。