Long noncoding RNA GAS5 inhibits progression of colorectal cancer by interacting with and triggering YAP phosphorylation and degradation and is negatively regulated by the m6A reader YTHDF3
Long noncoding RNA GAS5 inhibits progression of colorectal cancer by interacting with and triggering YAP phosphorylation and degradation and is negatively regulated by the m6A reader YTHDF3
复制标题
长非编码 RNA GAS5 通过与 YAP 相互作用并触发 YAP 磷酸化和降解来抑制结直肠癌的进展，并受到 m(6)A 阅读器 YTHDF3 的负调节。
DOI：
10.1186/s12943-019-1079-y
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发表时间：
2019-10-16
期刊：
影响因子：
37.3
通讯作者：
Li, Jianming
中科院分区：
文献类型：
--
作者：
Ni, Wen;Yao, Su;Li, Jianming
背景YAP 激活对于包括结直肠癌 (CRC) 在内的癌症发展至关重要。然而，目前尚不清楚 N6-甲基腺苷 (m(6)A) 修饰的长非编码 RNA (lncRNA) 转录物是否可以调节癌症进展中的 YAP 激活。我们研究了 lncRNA 与 YAP 信号传导和 CRC 进展中 m(6)A 修饰之间的功能联系。方法通过 RIP 测序、RNA FISH 和免疫荧光共染色测定筛选 YAP 相互作用的 lncRNA。通过生化方法研究了YAP和lncRNA GAS5之间的相互作用。 MeRIP 测序结合 lncRNA 测序用于鉴定 CRC 中 YTHDF3 的 m(6)A 修饰靶点。通过功能获得和功能丧失分析来测量 GAS5-YAP-YTHDF3 轴在体外和体内 CRC 进展中的功能。结果GAS5 直接与 YAP 的 WW 结构域相互作用，促进内源性 YAP 从细胞核易位到细胞质，促进磷酸化，随后泛素介导的 YAP 降解以抑制 CRC 体外和体内进展。值得注意的是，我们证明 m(6)A 阅读器 YTHDF3 不仅是 YAP 的新靶标，而且通过促进 m(6)A 修饰的 lncRNA GAS5 降解而成为 YAP 信号传导的关键参与者，这为 CRC 进展提供了新的见解。临床上，lncRNA GAS5 表达与 CRC 患者肿瘤中 YAP 和 YTHDF3 蛋白水平呈负相关。结论我们的研究揭示了 lncRNA GAS5-YAP-YTHDF3 轴的负功能环，并确定了结直肠癌进展中 m(6)A 诱导 YAP 信号传导中 GAS5 衰减的新机制，这可能为 CRC 治疗提供一种有前景的方法。
BackgroundYAP activation is crucial for cancer development including colorectal cancer (CRC). Nevertheless, it remains unclear whether N6-Methyladenosine (m(6)A) modified transcripts of long noncoding RNAs (lncRNAs) can regulate YAP activation in cancer progression. We investigated the functional link between lncRNAs and the m(6)A modification in YAP signaling and CRC progression.MethodsYAP interacting lncRNAs were screened by RIP-sequencing, RNA FISH and immunofluorescence co-staining assays. Interaction between YAP and lncRNA GAS5 was studied by biochemical methods. MeRIP-sequencing combined with lncRNA-sequencing were used to identify the m(6)A modified targets of YTHDF3 in CRC. Gain-of-function and Loss-of-function analysis were performed to measure the function of GAS5-YAP-YTHDF3 axis in CRC progression in vitro and in vivo.ResultsGAS5 directly interacts with WW domain of YAP to facilitate translocation of endogenous YAP from the nucleus to the cytoplasm and promotes phosphorylation and subsequently ubiquitin-mediated degradation of YAP to inhibit CRC progression in vitro and in vivo. Notably, we demonstrate the m(6)A reader YTHDF3 not only a novel target of YAP but also a key player in YAP signaling by facilitating m(6)A-modified lncRNA GAS5 degradation, which profile a new insight into CRC progression. Clinically, lncRNA GAS5 expressions is negatively correlated with YAP and YTHDF3 protein levels in tumors from CRC patients.ConclusionsOur study uncovers a negative functional loop of lncRNA GAS5-YAP-YTHDF3 axis, and identifies a new mechanism for m(6)A-induced decay of GAS5 on YAP signaling in progression of CRC which may offer a promising approach for CRC treatment.