基础医学与临床 ›› 2021, Vol. 41 ›› Issue (2): 159-164.

• 研究论文 • 上一篇    下一篇

C1orf194基因Ile122Asn突变致腓骨肌萎缩症的病理机制

陈诗恺, 陈韭铭, 孙顺昌*   

  1. 上海交通大学医学院附属瑞金医院北院 检验科, 上海 201801
  • 收稿日期:2020-05-06 修回日期:2020-06-28 出版日期:2021-02-05 发布日期:2021-01-19
  • 通讯作者: *shunchangsun@aliyun.com
  • 基金资助:
    国家自然科学基金(31571294)

Pathological mechanism of Charcot-Marie-Tooth disease caused by the Ile122Asn mutation in the C1orf194 gene

CHEN Shi-kai, CHEN Jiu-ming, SUN Shun-chang*   

  1. Department of Clinical Laboratory, Ruijin Hospital North, Shanghai Jiao Tong University School of Medicine, Shanghai 201801, China
  • Received:2020-05-06 Revised:2020-06-28 Online:2021-02-05 Published:2021-01-19
  • Contact: *shunchangsun@aliyun.com

摘要: 目的 初步探讨C1orf194基因内Ile122Asn突变导致常染色体显性遗传腓骨肌萎缩症(CMT)的分子机制。方法 通过PCR克隆构建人野生C1orf194基因和携带Ile122Asn突变的C1orf194基因,分别导入人神经母细胞瘤(SH-SY5Y)细胞内进行表达,通过稳定同位素标记氨基酸免疫沉淀分析和液相色谱确定候选相互作用蛋白,通过免疫印迹分别确定与野生c1orf194蛋白或突变c1orf194蛋白相互作用的蛋白。结果 在SH-SY5Y细胞内野生c1orf194蛋白与TBA1C、AACT及HSPA4L等3种蛋白存在相互作用,携带Ile122Asn突变的c1orf194蛋白则失去与这3种蛋白的相互作用。结论 C1orf194基因内的Ile122Asn突变影响c1orf194蛋白与相互作用蛋白的结合,或许影响其调控的分子路径,使患者表现为常染色体显性遗传腓骨肌萎缩症,这对进一步研究c1orf194蛋白功能有重要意义。

关键词: 腓骨肌萎缩症, C1orf194基因, 突变, 相互作用蛋白

Abstract: Objective To explore the molecular mechanism of the autosomal dominant Charcot-Marie-Tooth disease caused by the Ile122Asn mutation in the C1orf194 gene. Methods Human wild C1orf194 and Ile122Asn mutation C1orf194 genes were constructed by PCR cloning, and then expressed in SH-SY5Y cells. Candidate c1orf194-interacting proteins were screened by stable isotope labeling with amino acids in cell culture-immunoprecipitation following liquid chromatograph mass spectrometer/mass spectrometer operation. The c1orf194-interacting proteins were validated by Western blot. Results Wild c1orf194 interacts with TBA1C, AACT, and HSPA4L in SH-SY5Y cells. However, the Ile122Asn mutant c1orf194 failed to bind to them. Conclusions The results indicate that the C1orf194 gene mutation may lead to an autosomal dominant CMT by losing protein-protein interactions which are involved in different functional pathways. This conclusion may make contribution to clear understanding of the c1orf194 function.

Key words: Charcot-Marie-Tooth disease, C1orf194 gene, mutation, interacting protein

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