Maroney P A, Hannon G J, Denker J A, Nilsen T W
Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, OH 44106.
EMBO J. 1990 Nov;9(11):3667-73. doi: 10.1002/j.1460-2075.1990.tb07578.x.
The trans-spliced leader RNA (SL RNA) of nematodes resembles U snRNAs both in cap structure and in the presence of a consensus Sm binding site. We show here that synthetic SL RNA, synthesized by in vitro transcription, is efficiently used as a spliced leader donor in trans-splicing reactions catalyzed by a cell free extract prepared from developing embryos of the parasitic nematode, Ascaris lumbricoides. Efficient utilization of synthetic SL RNA requires a functional Sm binding site. Mutations within the Sm binding sequence that prevent immunoprecipitation by Sm antisera and prevent cap trimethylation abolish trans-splicing. The effect on trans-splicing is not due to undermethylation of the cap structure.
线虫的反式剪接前导RNA(SL RNA)在帽结构和存在共有Sm结合位点方面类似于U snRNA。我们在此表明,通过体外转录合成的合成SL RNA,在由寄生线虫蛔虫发育胚胎制备的无细胞提取物催化的反式剪接反应中,被有效地用作剪接前导供体。合成SL RNA的有效利用需要一个功能性的Sm结合位点。Sm结合序列内的突变会阻止Sm抗血清的免疫沉淀并阻止帽三甲基化,从而消除反式剪接。对反式剪接的影响并非由于帽结构的甲基化不足。