Department of Medicine, Division of Hematology/Oncology, Shands Cancer Center, University of Florida, Gainesville, FL, USA.
Cell Cycle. 2011 Jul 15;10(14):2390-9. doi: 10.4161/cc.10.14.16587.
We discovered and reported JAZ as a unique dsRNA binding zinc finger protein that functions as a direct, positive regulator of p53 transcriptional activity to mediate G1 cell cycle arrest in a mechanism involving upregulation of the p53 target gene, p21. We now find that JAZ can also negatively regulate the cell cycle in a novel, p53-independent mechanism resulting from the direct interaction with E2F1, a key intermediate in regulating cell proliferation and tumor suppression. JAZ associates with E2F1's central DNA binding/dimerization region and its C-terminal transactivation domain. Functionally, JAZ represses E2F1 transcriptional activity in association with repression of cyclin A expression and inhibition of G1/S transition. This mechanism involves JAZ-mediated inhibition of E2F1's specific DNA binding activity. JAZ directly binds E2F1 in vitro in a dsRNA-independent manner, and JAZ's dsRNA binding ZF domains, which are necessary for localizing JAZ to the nucleus, are required for repression of transcriptional activity in vivo. Importantly for specificity, siRNA-mediated "knockdown" of endogenous JAZ increases E2F transcriptional activity and releases cells from G1 arrest, indicating a necessary role for JAZ in this transition. Although JAZ can directly inhibit E2F1 activity independently of p53, if functional p53 is expressed, JAZ may exert a more potent inhibition of cell cycle following growth factor withdrawal. Therefore, JAZ plays a dual role in cell cycle regulation by both repressing E2F1 transcriptional activity and activating p53 to facilitate efficient growth arrest in response to cellular stress, which may potentially be exploited therapeutically for tumor growth inhibition.
我们发现并报道了 JAZ 是一种独特的 dsRNA 结合锌指蛋白,它作为 p53 转录活性的直接正向调节剂,通过上调 p53 靶基因 p21 来介导 G1 细胞周期停滞。我们现在发现,JAZ 还可以通过与 E2F1 的直接相互作用,以一种新的、不依赖 p53 的机制负调控细胞周期,E2F1 是调节细胞增殖和肿瘤抑制的关键中间物。JAZ 与 E2F1 的中央 DNA 结合/二聚化区域及其 C 端转录激活结构域相互作用。功能上,JAZ 通过抑制 cyclin A 的表达和抑制 G1/S 过渡来抑制 E2F1 的转录活性。这种机制涉及 JAZ 介导的 E2F1 特定 DNA 结合活性的抑制。JAZ 在体外以 dsRNA 非依赖的方式直接与 E2F1 结合,并且 JAZ 的 dsRNA 结合 ZF 结构域对于将 JAZ 定位到核内是必需的,对于体内转录活性的抑制也是必需的。对于特异性很重要的是,siRNA 介导的内源性 JAZ“敲低”增加了 E2F 的转录活性并使细胞从 G1 期停滞中释放出来,这表明 JAZ 在这种转变中起着必要的作用。尽管 JAZ 可以独立于 p53 直接抑制 E2F1 的活性,但如果有功能性的 p53 表达,JAZ 在生长因子撤出后可能会对细胞周期产生更强的抑制作用。因此,JAZ 通过抑制 E2F1 的转录活性和激活 p53 来发挥双重作用,以促进细胞应激时的有效生长停滞,这可能在肿瘤生长抑制的治疗中具有潜在的应用价值。