Expression of human FUS/TLS in yeast leads to protein aggregation and cytotoxicity, recapitulating key features of FUS proteinopathy.
Expression of human FUS/TLS in yeast leads to protein aggregation and cytotoxicity, recapitulating key features of FUS proteinopathy.
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DOI：
10.1007/s13238-011-1014-5
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发表时间：
2011-02
期刊：
影响因子：
21.1
通讯作者：
Wu JY
中科院分区：
文献类型：
--
作者：
Fushimi K;Long C;Jayaram N;Chen X;Li L;Wu JY
肉瘤融合/脂肪肉瘤易位（FUS/TLS）基因的突变与肌萎缩侧索硬化症（ALS）有关。FUS阳性神经病理学在一系列神经退行性疾病中有报道，包括ALS和额颞叶变性伴泛素阳性病理学（FTLDU）。为了检测蛋白质聚集和细胞毒性，我们在酵母中表达了人FUS蛋白。野生型或ALS相关的R524 S或P525 L突变体FUS在酵母细胞中的表达导致聚集体和细胞毒性的形成，其中两种ALS突变体显示出增加的细胞毒性。因此，表达人FUS蛋白的酵母细胞概括了FUS阳性神经退行性疾病的关键特征。有趣的是，用碘化丙啶染色的表达FUS的酵母细胞中有很大一部分没有可检测到的蛋白质聚集体，这表明FUS表达引起的膜损伤和细胞损伤可能在蛋白质聚集体变得可通过显微镜检测到之前发生，并且聚集体的形成可能会保护细胞免受FUS介导的细胞毒性。FUS的N-末端含有QGSY和G富集区，足以形成聚集体，但不具有细胞毒性。C-末端结构域，其中包含一簇突变，没有显示出聚集或细胞毒性。与TDP-43在酵母中表达时类似，FUS蛋白具有在不存在其他人蛋白质的情况下形成聚集体的固有性质。另一方面，由FUS形成的聚集体是硫磺素T阳性的，并且对0.5%的肌氨酰具有抗性，这与在酵母细胞中表达时的TDP-43不同。此外，SDP-43和FUS在聚集体形成和细胞毒性方面表现出不同的结构域要求。
Mutations in the fused in sarcoma/translocated in liposarcoma (FUS/TLS) gene have been associated with amyotrophic lateral sclerosis (ALS). FUS-positive neuro-pathology is reported in a range of neurodegenerative diseases, including ALS and fronto-temporal lobar degeneration with ubiquitin-positive pathology (FTLDU). To examine protein aggregation and cytotoxicity, we expressed human FUS protein in yeast. Expression of either wild type or ALS-associated R524S or P525L mutant FUS in yeast cells led to formation of aggregates and cytotoxicity, with the two ALS mutants showing increased cytotoxicity. Therefore, yeast cells expressing human FUS protein recapitulate key features of FUS-positive neurodegenerative diseases. Interestingly, a significant fraction of FUS expressing yeast cells stained by propidium iodide were without detectable protein aggregates, suggesting that membrane impairment and cellular damage caused by FUS expression may occur before protein aggregates become microscopically detectable and that aggregate formation might protect cells from FUS-mediated cytotoxicity. The N-terminus of FUS, containing the QGSY and G rich regions, is sufficient for the formation of aggregates but not cytotoxicity. The C-terminal domain, which contains a cluster of mutations, did not show aggregation or cytotoxicity. Similar to TDP-43 when expressed in yeast, FUS protein has the intrinsic property of forming aggregates in the absence of other human proteins. On the other hand, the aggregates formed by FUS are thioflavin T-positive and resistant to 0.5% sarkosyl, unlike TDP-43 when expressed in yeast cells. Furthermore, TDP-43 and FUS display distinct domain requirements in aggregate formation and cytotoxicity.
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影响因子：
12.7
作者：
Urwin H;Josephs KA;Rohrer JD;Mackenzie IR;Neumann M;Authier A;Seelaar H;Van Swieten JC;Brown JM;Johannsen P;Nielsen JE;Holm IE;FReJA Consortium;Dickson DW;Rademakers R;Graff-Radford NR;Parisi JE;Petersen RC;Hatanpaa KJ;White CL 3rd;Weiner MF;Geser F;Van Deerlin VM;Trojanowski JQ;Miller BL;Seeley WW;van der Zee J;Kumar-Singh S;Engelborghs S;De Deyn PP;Van Broeckhoven C;Bigio EH;Deng HX;Halliday GM;Kril JJ;Munoz DG;Mann DM;Pickering-Brown SM;Doodeman V;Adamson G;Ghazi-Noori S;Fisher EM;Holton JL;Revesz T;Rossor MN;Collinge J;Mead S;Isaacs AM
通讯作者：
Isaacs AM
影响因子：
56.9
作者：
Kwiatkowski, T. J., Jr.;Bosco, D. A.;Brown, R. H., Jr.
通讯作者：
Brown, R. H., Jr.
影响因子：
14.5
作者：
Neumann, Manuela;Rademakers, Rosa;Mackenzie, Ian R. A.
通讯作者：
Mackenzie, Ian R. A.
影响因子：
4.8
作者：
Johnson, Brian S.;Snead, David;Gitler, Aaron D.
通讯作者：
Gitler, Aaron D.
DOI：
10.1111/j.1750-3639.2010.00413.x
发表时间：
2010-11
期刊：
Brain pathology (Zurich, Switzerland)
影响因子：
--
作者：
Huang EJ;Zhang J;Geser F;Trojanowski JQ;Strober JB;Dickson DW;Brown RH Jr;Shapiro BE;Lomen-Hoerth C
通讯作者：
Lomen-Hoerth C