Dunstan H M, Young L S, Sprague K U
Institute of Molecular Biology, University of Oregon, Eugene 97403.
Mol Cell Biol. 1994 Jun;14(6):3588-95. doi: 10.1128/mcb.14.6.3588-3595.1994.
Promoter-specific transcription by silkworm RNA polymerase III is dependent on several transcription factors (TFs) in addition to the polymerase itself. The activities present in silk gland nuclear extracts that are necessary to reconstitute transcription from class III genes in vitro have been resolved into several partially purified components. These include TFIIIR, which is unusual because it is composed of RNA. Here, we identify the RNA that provides TFIIIR activity as silkworm tRNA(IleIAU). This conclusion is based on copurification of tRNA(IleIAU) with TFIIIR activity, TFIIIR activity in synthetic tRNA(Ile), and hybrid selection of TFIIIR activity by antisense tRNA(IleIAU). We have tested the ability of a variety of other tRNAs to stimulate transcription and find that TFIIIR activity is highly specific to silkworm tRNA(IleIAU).
家蚕RNA聚合酶III的启动子特异性转录除了依赖于聚合酶本身外,还依赖于几种转录因子(TFs)。丝腺核提取物中存在的、在体外重建III类基因转录所必需的活性已被解析为几个部分纯化的组分。其中包括TFIIIR,它不同寻常之处在于它由RNA组成。在这里,我们鉴定出提供TFIIIR活性的RNA为家蚕tRNA(IleIAU)。这一结论基于tRNA(IleIAU)与TFIIIR活性的共纯化、合成tRNA(Ile)中的TFIIIR活性以及反义tRNA(IleIAU)对TFIIIR活性的杂交筛选。我们测试了多种其他tRNA刺激转录的能力,发现TFIIIR活性对家蚕tRNA(IleIAU)具有高度特异性。