金振辉, 范礼斌, 姜天霞, 邱小波. 蛋白酶体泛素受体亚基Rpn13转录抑制调控机制探究[J]. 北京师范大学学报(自然科学版). DOI: 10.12202/j.0476-0301.2024053
引用本文: 金振辉, 范礼斌, 姜天霞, 邱小波. 蛋白酶体泛素受体亚基Rpn13转录抑制调控机制探究[J]. 北京师范大学学报(自然科学版). DOI: 10.12202/j.0476-0301.2024053
JIN Zhenhui, FAN Libin, JIANG Tianxia, QIU Xiaobo. Regulatorymechanismoftranscriptionalrepressionof the proteasomal ubiquitin receptor Rpn13[J]. Journal of Beijing Normal University(Natural Science). DOI: 10.12202/j.0476-0301.2024053
Citation: JIN Zhenhui, FAN Libin, JIANG Tianxia, QIU Xiaobo. Regulatorymechanismoftranscriptionalrepressionof the proteasomal ubiquitin receptor Rpn13[J]. Journal of Beijing Normal University(Natural Science). DOI: 10.12202/j.0476-0301.2024053

蛋白酶体泛素受体亚基Rpn13转录抑制调控机制探究

Regulatorymechanismoftranscriptionalrepressionof the proteasomal ubiquitin receptor Rpn13

  • 摘要: 摘要哺乳动物Rpn13作为26S蛋白酶体泛素受体亚基,其N端结合泛素化底物,C端结合于去泛素化酶UCH37,介导大部分蛋白质通过泛素-蛋白酶体通路降解,参与调控细胞自噬、神经元功能和生殖细胞发育等.蛋白酶体抑制剂是肿瘤防治的重要措施,且靶向Rpn13能够克服蛋白酶体抑制剂的耐药性 然而,Rpn13基因转录抑制调控机制仍不清楚.本文通过基因表达谱交互式分析发现,Rpn13基因在结肠癌、淋巴癌、胰腺癌、直肠癌、胃癌以及胸腺癌等中高表达.双荧光素酶报告实验显示Rpn13启动子区域−454~58 bp的活性较高,为其转录调控的主要区域.Rpn13启动子序列中存在多个转录因子的结合位点,如CTCFL(CCCTC binding factor-like)、CTCF(CCCTC binding factor)、NRF1(Nuclear respiratory factor 1)和FOXO3(Forkhead box O3)等,并且它们均会抑制Rpn13启动子活性.进一步的突变实验显示CTCFL结合在启动−63 bp~−52 bp序列.本研究不仅发现Rpn13在癌组织中高表达,也初步揭示其转录抑制机制,为探究Rpn13在肿瘤发生发展中的作用机制提供线索.

     

    Abstract: Mammalian Rpn13, as a 26S proteasome ubiquitin receptor subunit, binds to the ubiquitin substrate at the N terminal and deubiquitin enzyme UCH37 at the C end, mediates the degradation of most proteins through the ubiquitin-proteasome pathway, and is involved in the regulation of autophagy, neuron function and germ cell development.Proteasome inhibitors are important measures for tumor prevention and treatment, and targeting Rpn13 can overcome the resistance of proteasome inhibitors. However, the transcriptional repression mechanism of its gene expression is still unclear. Through the interactive analysis of gene expression profile, it was found that Rpn13 gene was highly expressed in many cancer types, especially in colon, lymphoma, pancreatic, rectal, gastric and thoracic cancers. The activity of the regulatory region -454~-58bp of the Rpn13 promoter was found to be high by the double luciferase report assay. Binding sites of multiple transcription factors, includingCTCFL(CCCTC binding factor-like),CTCF(CCCTC binding factor),NRF1(Nuclear respiratory factor 1)andFOXO3 (Forkhead box O3),were predicted on the promoter sequence of Rpn13. We constructed expression plasmids of these transcription factors, and found that they all inhibited the activity of the Rpn13 promoter. By further mutating the corresponding transcription factor binding sites of the Rpn13 promoter, CTCFL was found to bind the -63bp~-52bp sequence of the Rpn13 promoter. These studies not only analyzed the expression of Rpn13 gene in cancer tissues, but also revealed the possible transcription inhibition mechanism of Rpn13, providing useful clues for exploring the mechanism of Rpn13 in tumor occurrence and development.

     

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