SUMO1 modification of KHSRP regulates tumorigenesis by preventing the TL-G-Rich miRNA biogenesis.
SUMO1 modification of KHSRP regulates tumorigenesis by preventing the TL-G-Rich miRNA biogenesis.
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KHSRP 的 SUMO1 修饰通过阻止富含 TL-G 的 miRNA 生物发生来调节肿瘤发生
DOI：
10.1186/s12943-017-0724-6
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发表时间：
2017-10-11
期刊：
影响因子：
37.3
通讯作者：
Yu J
中科院分区：
文献类型：
--
作者：
Yuan H;Deng R;Zhao X;Chen R;Hou G;Zhang H;Wang Y;Xu M;Jiang B;Yu J
microRNA（miRNAs）是参与包括癌症在内的多种生理和病理过程的重要调节因子。SUMO（small ubiquitin-like modifier）是一种可逆的蛋白质修饰剂。我们最近发现TARBP2和DGCR8的SUMO化参与了miRNA通路的调节。KHSRP是一种单链核酸结合蛋白，在转录和mRNA降解中起作用，也是Drosha-DGCR8复合物的一个组分，促进miRNA的生物合成。使用Ni2 +-NTA亲和下拉或免疫沉淀（IP）的体内SUMO化测定和基于大肠杆菌的体外SUMO化测定来分析KHSRP的SUMO化。采用细胞核/胞浆分级分离法和免疫荧光染色法观察KHSRP的定位。采用高通量miRNA测序、定量RT-PCR和RNA免疫沉淀分析（RIP）等方法研究KHSRP SUMO 1修饰对miRNA生物合成的影响。通过软琼脂集落形成、迁移、血管生成拟态（VM）和三维细胞培养等方法检测肿瘤细胞的表型，并通过小鼠异种移植瘤模型验证SUMO 1修饰KHSRP对体内肿瘤发生的调控作用。SUMO 1修饰KHSRP的主要位点K87，这种修饰在微环境缺氧时可以增加，而用生长因子处理可以减少。KHSRP的SUMO 1修饰抑制其与pri-miRNA/Drosha-DGCR 8复合物的相互作用，并可能增加其从细胞核到细胞质的易位。因此，KHSRP的SUMO 1修饰损害了从pri-miRNA加工pre-miRNA的步骤，pri-miRNA特别是在其末端环（TL）中具有短的富含G的片段，导致TL-G-Rich miRNA的子集如let-7家族的下调和随后的肿瘤发生。我们的数据证明了miRNA生物发生途径如何通过KHSRP的可逆SUMO 1修饰与肿瘤发生和癌症进展相关。
MicroRNAs (miRNAs) are important regulators involved in diverse physiological and pathological processes including cancer. SUMO (small ubiquitin-like modifier) is a reversible protein modifier. We recently found that SUMOylation of TARBP2 and DGCR8 is involved in the regulation of the miRNA pathway. KHSRP is a single stranded nucleic acid binding protein with roles in transcription and mRNA decay, and it is also a component of the Drosha-DGCR8 complex promoting the miRNA biogenesis. The in vivo SUMOylation assay using the Ni2+-NTA affinity pulldown or immunoprecipitation (IP) and the in vitro E.coli-based SUMOylation assay were used to analyze SUMOylation of KHSRP. Nuclear/Cytosol fractionation assay and immunofluorescent staining were used to observe the localization of KHSRP. High-throughput miRNA sequencing, quantantive RT-PCR and RNA immunoprecipitation assay (RIP) were employed to determine the effects of KHSRP SUMO1 modification on the miRNA biogenesis. The soft-agar colony formation, migration, vasculogenic mimicry (VM) and three-dimensional (3D) cell culture assays were performed to detect the phenotypes of tumor cells in vitro, and the xenograft tumor model in mice was conducted to verify that SUMO1 modification of KHSRP regulated tumorigenesis in vivo. KHSRP is modified by SUMO1 at the major site K87, and this modification can be increased upon the microenvironmental hypoxia while reduced by the treatment with growth factors. SUMO1 modification of KHSRP inhibits its interaction with the pri-miRNA/Drosha-DGCR8 complex and probably increases its translocation from the nucleus to the cytoplasm. Consequently, SUMO1 modification of KHSRP impairs the processing step of pre-miRNAs from pri-miRNAs which especially harbor short G-rich stretches in their terminal loops (TL), resulting in the downregulation of a subset of TL-G-Rich miRNAs such as let-7 family and consequential tumorigenesis. Our data demonstrate how the miRNA biogenesis pathway is connected to tumorigenesis and cancer progression through the reversible SUMO1 modification of KHSRP.
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影响因子：
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作者：
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通讯作者：
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影响因子：
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