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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >ALS-associated mutations in TDP-43 increase its stability and promote TDP-43 complexes with FUS/TLS
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ALS-associated mutations in TDP-43 increase its stability and promote TDP-43 complexes with FUS/TLS

机译:TDP-43中与ALS相关的突变增加了其稳定性并促进了FUS / TLS的TDP-43复合物

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摘要

Dominant mutations in two functionally related DNA/RNA-binding proteins, trans-activating response region (TAR) DNA-binding protein with a molecular mass of 43 KDa (TDP-43) and fused in sarcoma/ translocation in liposarcoma (FUS/TLS), cause an inherited form of ALS that is accompanied by nuclear and cytoplasmic aggregates containing TDP-43 or FUS/TLS. Using isogenic cell lines expressing wild-type or ALS-linked TDP-43 mutants and fibroblasts from a human patient, pulse-chase radiolabeling of newly synthesized proteins is used to determine, surprisingly, that ALS-linked TDP-43 mutant polypeptides are more stable than wild-type TDP-43. Tandem-affinity purification and quantitative mass spectrometry are used to identify TDP-43 complexes not only with heterogeneous nuclear ribonucleoproteins family proteins, as expected, but also with components of Drosha microprocessor complexes, consistent with roles for TDP-43 in both mRNA processing and microRNA biogenesis. A fraction of TDP-43 is shown to be complexed with FUS/TLS, an interaction substantially enhanced by TDP-43 mutants. Taken together, abnormal stability of mutant TDP-43 and its enhanced binding to normal FUS/TLS imply a convergence of pathogenic pathways from mutant TDP-43 and FUS/TLS in ALS.
机译:两个功能相关的DNA / RNA结合蛋白,分子量为43 KDa的反式激活反应区(TAR)DNA结合蛋白(TDP-43)和融合在肉瘤/脂肪肉瘤易位(FUS / TLS)中的显着突变引起遗传形式的ALS,并伴有含有TDP-43或FUS / TLS的核和细胞质聚集体。使用表达来自人类患者的野生型或ALS连接的TDP-43突变体和成纤维细胞的等基因细胞系,对新合成的蛋白质进行脉冲追踪放射性标记,令人惊讶地确定了ALS连接的TDP-43突变体多肽更稳定比野生型TDP-43串联亲和纯化和定量质谱用于鉴定TDP-43复合物,不仅与预期的异质核糖核蛋白家族蛋白一样,而且还与Drosha微处理器复合物的成分一起鉴定,这与TDP-43在mRNA加工和microRNA中的作用一致生物发生。 TDP-43的一部分显示与FUS / TLS复合,这种相互作用被TDP-43突变体大大增强。两者合计,突变体TDP-43的异常稳定性及其与正常FUS / TLS的增强结合暗示ALS中来自突变体TDP-43和FUS / TLS的致病性途径趋同。

著录项

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  • 作者单位

    Ludwig Institute for Cancer Research and Departments of University of California at San Diego, La Jolla, CA 92093-0670 Neuroscience and University of California at San Diego, La Jolla, CA 92093-0670 Cellular and Molecular Medicine, University of California at San Diego, La Jolla, CA 92093-0670;

    rnLudwig Institute for Cancer Research and Departments of University of California at San Diego, La Jolla, CA 92093-0670;

    rnLudwig Institute for Cancer Research and Departments of University of California at San Diego, La Jolla, CA 92093-0670 Cellular and Molecular Medicine, University of California at San Diego, La Jolla, CA 92093-0670;

    rnLudwig Institute for Cancer Research and Departments of University of California at San Diego, La Jolla, CA 92093-0670 Cellular and Molecular Medicine, University of California at San Diego, La Jolla, CA 92093-0670;

    rnLudwig Institute for Cancer Research and Departments of University of California at San Diego, La Jolla, CA 92093-0670;

    rnLudwig Institute for Cancer Research and Departments of University of California at San Diego, La Jolla, CA 92093-0670 Cellular and Molecular Medicine, University of California at San Diego, La Jolla, CA 92093-0670;

    rnLudwig Institute for Cancer Research and Departments of University of California at San Diego, La Jolla, CA 92093-0670 Neuroscience and University of California at San Diego, La Jolla, CA 92093-0670 Cellular and Molecular Medicine, University of California at San Diego, La Jolla, CA 92093-0670;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    mass spectrometry; protein stability; amyotrophic lateral sclerosis; microRNA; ribonucleoproteins;

    机译:质谱;蛋白质稳定性;肌萎缩性侧索硬化症;微小RNA;核糖核蛋白;

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