Maraia R J
Laboratory of Molecular Growth Regulation, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
Nucleic Acids Res. 1991 Oct 25;19(20):5695-702. doi: 10.1093/nar/19.20.5695.
B1 (Alu-equivalent) is a murine short interspersed element whose amplification probably involved an RNA intermediate. B1-homologous RNA comprise a population of heterogenous transcripts of questionable function. A cloned B1 is expressed in the injected frog oocyte by RNA polymerase III transcription, ribonucleoprotein formation, post-transcriptional 3'-processing, and nucleocytoplasmic transport. The present study characterizes small cytoplasmic B1 transcripts of mouse cells. Analyses of ten cDNA clones revealed a subset of a high degree of sequence identity (98%) from which a novel consensus was developed. Structural analyses of these RNAs demonstrated a conserved Alu domain originally identified as part of the 7SL RNA within the translational control domain of the signal recognition particle, while this structure was not conserved in the majority of B1s in the sequence database. Furthermore, it was demonstrated that 3'-processing occurred in only a subset of B1 transcripts in-vitro using homologous nuclear extracts, and in the injected oocyte. The data demonstrate that a limited set of B1 sequences are expressed as processed RNA polymerase III-transcripts of a high degree of structural conservation. Although this subset is transcriptionally active, the selective expression may be due to regulation at the levels of processing and cytoplasmic accumulation. Their lack of Poly-(A) or 3'-oligo-(U) tracts argue that these RNAs are unlikely to represent transposition intermediates. Rather, their cytosolic compartmentalization and conservation of a biologically recognized structure, suggests potential involvement in other aspects of cellular metabolism.
B1(Alu 等效元件)是一种小鼠短散在元件,其扩增可能涉及 RNA 中间体。B1 同源 RNA 由一群功能存疑的异质转录本组成。通过 RNA 聚合酶 III 转录、核糖核蛋白形成、转录后 3' 加工和核质运输,克隆的 B1 在注射的蛙卵母细胞中表达。本研究对小鼠细胞中的小细胞质 B1 转录本进行了表征。对十个 cDNA 克隆的分析揭示了一个高度序列同一性(98%)的子集,并据此形成了一个新的共有序列。对这些 RNA 的结构分析表明,在信号识别颗粒的翻译控制域内,存在一个最初被鉴定为 7SL RNA 一部分的保守 Alu 结构域,而在序列数据库中的大多数 B1 中,这种结构并不保守。此外,使用同源核提取物在体外以及在注射的卵母细胞中证实,3' 加工仅发生在 B1 转录本的一个子集中。数据表明,一组有限的 B1 序列被表达为具有高度结构保守性的经加工的 RNA 聚合酶 III 转录本。尽管这个子集具有转录活性,但选择性表达可能是由于加工和细胞质积累水平的调控。它们缺乏 Poly-(A) 或 3'-oligo-(U) 序列,这表明这些 RNA 不太可能代表转座中间体。相反,它们在细胞质中的区室化以及生物识别结构的保守性,表明它们可能参与细胞代谢的其他方面。