ALS-associated mutations in TDP-43 increase its stability and promote TDP-43 complexes with FUS/TLS
ALS-associated mutations in TDP-43 increase its stability and promote TDP-43 complexes with FUS/TLS
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DOI：
10.1073/pnas.1008227107
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发表时间：
2010-07-27
影响因子：
11.1
通讯作者：
Cleveland, Don W.
中科院分区：
文献类型：
--
作者：
Ling, Shuo-Chien;Albuquerque, Claudio P.;Cleveland, Don W.
在两个功能相关的DNA/RNA结合蛋白中的显着突变，分子质量为43 kDa（TDP-43）的反式激活反应区域（TAR）DNA结合蛋白，并在Liposarcoma（FUS/TLS）中的肉瘤/转运中融合，导致一种遗传形式的ALS，伴随着包含TDP-43或的核和细胞质聚集体FUS/TLS。使用表达野生型或ALS连接的TDP-43突变体和来自人类患者的成纤维细胞的等生细胞系，新合成蛋白的脉冲练习式放射性标记用于确定ALS链接的TDP-43突变肽是更稳定的稳定性的。比野生型TDP-43。如预期生物发生。 TDP-43的一部分被证明与FUS/TLS复合了，TDP-43突变体大大增强了这种相互作用。两者合计，突变体TDP-43的异常稳定性及其与正常FUS/TLS的结合意味着ALS中突变体TDP-43和FUS/TLS的致病途径收敛。
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.