From the Abramson Family Cancer Research Institute, Department of Cancer Biology, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania 19104.
J Biol Chem. 2014 Apr 4;289(14):9902-8. doi: 10.1074/jbc.M113.520692. Epub 2014 Feb 21.
Multiple endocrine neoplasia type I (MEN1) is an inherited syndrome that includes susceptibility to pancreatic islet hyperplasia. This syndrome results from mutations in the MEN1 gene, which encodes menin protein. Menin interacts with several transcription factors, including JunD, and inhibits their activities. However, the precise mechanism by which menin suppresses gene expression is not well understood. Here, we show that menin interacts with arsenite-resistant protein 2 (ARS2), a component of the nuclear RNA CAP-binding complex that is crucial for biogenesis of certain miRNAs including let-7a. The levels of primary-let-7a (pri-let-7a) are not affected by menin; however, the levels of mature let-7a are substantially decreased upon Men1 excision. Let-7a targets, including Insr and Irs2, pro-proliferative genes that are crucial for insulin-mediated signaling, are up-regulated in Men1-excised cells. Inhibition of let-7a using anti-miRNA in wild type cells is sufficient to enhance the expression of insulin receptor substrate 2 (IRS2) to levels observed in Men1-excised cells. Depletion of menin does not affect the expression of Drosha and CBP80, but substantially impairs the processing of pri-miRNA to pre-miRNA. Ars2 knockdown decreased let-7a processing in menin-expressing cells but had little impact on let-7a levels in menin-excised cells. As IRS2 is known to mediate insulin signaling and insulin/mitogen-induced cell proliferation, these findings collectively unravel a novel mechanism whereby menin suppresses cell proliferation, at least partly by promoting the processing of certain miRNAs, including let-7a, leading to suppression of Irs2 expression and insulin signaling.
多发性内分泌肿瘤 1 型(MEN1)是一种遗传性综合征,包括胰岛增生易感性。该综合征是由 MEN1 基因突变引起的,该基因编码 menin 蛋白。Menin 与几种转录因子相互作用,包括 JunD,并抑制它们的活性。然而,menin 抑制基因表达的确切机制尚不清楚。在这里,我们表明 menin 与砷酸盐抗性蛋白 2(ARS2)相互作用,ARS2 是核 RNA CAP 结合复合物的一个组成部分,对于某些 miRNA(包括 let-7a)的生物发生至关重要。menin 不会影响初级 let-7a(pri-let-7a)的水平;然而,成熟 let-7a 的水平在 Men1 缺失后显著降低。let-7a 的靶标,包括 Insr 和 Irs2,是胰岛素介导的信号传导所必需的促增殖基因,在 Men1 缺失的细胞中上调。在野生型细胞中使用抗 miRNA 抑制 let-7a 足以将胰岛素受体底物 2(IRS2)的表达水平提高到 Men1 缺失细胞中观察到的水平。Menin 的缺失不影响 Drosha 和 CBP80 的表达,但会严重影响 pri-miRNA 向 pre-miRNA 的加工。Ars2 的敲低降低了 menin 表达细胞中的 let-7a 加工,但对 menin 缺失细胞中的 let-7a 水平几乎没有影响。由于 IRS2 已知介导胰岛素信号和胰岛素/有丝分裂原诱导的细胞增殖,这些发现共同揭示了一种新的机制,即 menin 通过促进某些 miRNA(包括 let-7a)的加工来抑制细胞增殖,至少部分地抑制 IRS2 表达和胰岛素信号。