Kobayashi S, Agui K, Kamo S, Li Y, Anzai K
Department of Biochemistry, College of Pharmacy, Nihon University, Narashinodai, Funabashi, Chiba, 274-8555, Japan.
Biochem Biophys Res Commun. 2000 Oct 22;277(2):341-7. doi: 10.1006/bbrc.2000.3683.
BC1 RNA is preferentially expressed in neural cells by RNA polymerase III (Pol III) and forms ribonucleoprotein particles (RNP) in the somatodendritic domain of neurons. Our previous studies have suggested that, in the nucleus, BC1 RNA forms an RNP containing a nuclear protein(s) that participates in the transcription of the BC1 RNA gene. In this study, we have shown that newly synthesized BC1 RNA in purified brain nuclear extracts is immunoprecipitated by an antibody against Pur alpha. Pur alpha is a protein that binds single-stranded DNA and RNA and is known to regulate transcription of Pol II system. Although BC1 RNA is transcribed by Pol III, the BC1 RNA gene has two putative Pur alpha binding sites, which Pur alpha specifically recognizes. Point mutations within these sites reduced transcriptional activity in vitro. Furthermore, transcription was inhibited by depletion of Pur alpha from the nuclear extracts, either by the coexistence of its binding region of BC1 RNA or by the antibody that was able to precipitate the nuclear BC1 RNP. These observations suggest that BC1 RNA associates with Pur alpha which is involved in the transcription of the BC1 RNA gene.
BC1 RNA 由 RNA 聚合酶 III(Pol III)在神经细胞中优先表达,并在神经元的树突体区域形成核糖核蛋白颗粒(RNP)。我们之前的研究表明,在细胞核中,BC1 RNA 形成一种包含参与 BC1 RNA 基因转录的核蛋白的 RNP。在本研究中,我们发现纯化的脑细胞核提取物中新合成的 BC1 RNA 可被抗 Pur alpha 的抗体免疫沉淀。Pur alpha 是一种结合单链 DNA 和 RNA 的蛋白质,已知其可调节 Pol II 系统的转录。尽管 BC1 RNA 由 Pol III 转录,但 BC1 RNA 基因有两个假定的 Pur alpha 结合位点,Pur alpha 可特异性识别这些位点。这些位点内的点突变降低了体外转录活性。此外,通过 BC1 RNA 结合区域的共存或能够沉淀核 BC1 RNP 的抗体从核提取物中耗尽 Pur alpha,转录受到抑制。这些观察结果表明,BC1 RNA 与参与 BC1 RNA 基因转录的 Pur alpha 相关联。