Murphy J T, Burgess R R, Dahlberg J E, Lund E
Cell. 1982 May;29(1):265-74. doi: 10.1016/0092-8674(82)90111-8.
Transcription of U1 small nuclear RNA from a 592 bp fragment of human DNA was analyzed in vivo and in vitro. When injected into Xenopus laevis oocyte nuclei, the cloned DNA is transcribed by RNA polymerase II to make human U1 snRNA. Thus the sequences of this fragment are sufficient for expression of the U1 snRNA gene. Moreover, injection of templates carrying deletions of flanking sequences demonstrates that the DNA sequences required for in vivo transcription are located at least 100 nucleotides upstream from the point corresponding to the 5' end of mature U1 snRNA. In vitro transcription in a HeLa cell extract leads to synthesis not of mature U1 snRNA, but of a larger molecule starting 183 nucleotides upstream from the site corresponding to the 5' end of mature U1 snRNA. Transcription from this upstream promoter also is catalyzed by RNA polymerase II, and is comparable in efficiency with the very strong major late promoter of adenovirus 2. We propose that U1 snRNA is synthesized in vivo as a precursor that is processed by an enzyme or enzymes missing from our extracts.
对来自人类DNA的592 bp片段的U1小核RNA转录进行了体内和体外分析。当将克隆的DNA注射到非洲爪蟾卵母细胞核中时,它会被RNA聚合酶II转录以产生人类U1 snRNA。因此,该片段的序列足以用于U1 snRNA基因的表达。此外,注射携带侧翼序列缺失的模板表明,体内转录所需的DNA序列位于对应于成熟U1 snRNA 5'端的点上游至少100个核苷酸处。在HeLa细胞提取物中进行的体外转录导致合成的不是成熟的U1 snRNA,而是一个更大的分子,该分子从对应于成熟U1 snRNA 5'端的位点上游183个核苷酸处开始。来自该上游启动子的转录也由RNA聚合酶II催化,并且在效率上与腺病毒2的非常强的主要晚期启动子相当。我们提出,U1 snRNA在体内作为前体合成,该前体由我们提取物中缺失的一种或多种酶进行加工。