Single synonymous mutation in factor IX alters protein properties and underlies haemophilia B.
Single synonymous mutation in factor IX alters protein properties and underlies haemophilia B.
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DOI：
10.1136/jmedgenet-2016-104072
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发表时间：
2017-05
影响因子：
4
通讯作者：
Kimchi-Sarfaty C
中科院分区：
文献类型：
--
作者：
Simhadri VL;Hamasaki-Katagiri N;Lin BC;Hunt R;Jha S;Tseng SC;Wu A;Bentley AA;Zichel R;Lu Q;Zhu L;Freedberg DI;Monroe DM;Sauna ZE;Peters R;Komar AA;Kimchi-Sarfaty C
血友病B是由F9基因的遗传变异引起的。其中大多数是改变凝血因子IX（FIX）一级结构的非同义突变。然而，临床上有报道同义突变c.459G>A（缬氨酸107缬氨酸）可导致轻度血友病B（FIX凝血活性为正常的15 - 20%）。这些患者的F9 mRNA未显示内含子的跳跃或保留以及/或者mRNA水平的变化，这表明mRNA的完整性对受影响个体的疾病起源没有影响。本研究的目的是阐明能够解释具有这种同义突变的患者疾病表现的分子机制。
我们通过计算机分析以及在稳定细胞系中重现c.459G>A（缬氨酸107缬氨酸）突变来分析FIX缺乏的分子机制。利用构象敏感和非构象敏感抗体、有限的胰蛋白酶消化、FIX活性测定、与其他蛋白质的相互作用以及翻译后修饰来评估同义突变的生物物理和生物化学后果。
发现F9中的缬氨酸107缬氨酸同义突变显著降低了FIX的表达。我们的结果表明，该突变减缓了FIX的翻译并影响其构象，导致细胞外蛋白质水平降低。改变后的构象没有改变突变蛋白的比活性。
确定了与血友病B相关的F9基因中一个同义突变（缬氨酸107缬氨酸）的致病基础。对这些同义变异的机制理解为指导和开发未来的治疗方法提供了可能性。
Hemophilia B is caused by genetic aberrations in the F9 gene. The majority of these are non-synonymous mutations that alter the primary structure of blood coagulation Factor IX (FIX). However, a synonymous mutation c.459G>A (Val107Val) was clinically reported to result in mild hemophilia B (FIX coagulant activity 15–20% of normal). The F9 mRNA of these patients showed no skipping or retention of introns and/or change in mRNA levels, suggesting that mRNA integrity does not contribute to the origin of the disease in affected individuals. The aim of this study is to elucidate the molecular mechanisms that can explain disease manifestations in patients with this synonymous mutation. We analyze the molecular mechanisms underlying the FIX deficiency through in silico analysis and reproducing the c.459G>A (Val107Val) mutation in stable cell lines. Conformation and non-conformation sensitive antibodies, limited trypsin digestion, activity assays for FIX, interaction with other proteins, and post-translation modifications were used to evaluate the biophysical and biochemical consequences of the synonymous mutation. The Val107Val synonymous mutation in F9 was found to significantly diminish FIX expression. Our results suggest that this mutation slows FIX translation and affects its conformation resulting in decreased extracellular protein level. The altered conformation did not change the specific activity of the mutated protein. The pathogenic basis for one synonymous mutation (Val107Val) in the F9 gene associated with hemophilia B was determined. A mechanistic understanding of these synonymous variants yields potential for guiding and developing future therapeutic treatments.
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影响因子：
3.7
作者：
Edwards NC;Hing ZA;Perry A;Blaisdell A;Kopelman DB;Fathke R;Plum W;Newell J;Allen CE;S G;Shapiro A;Okunji C;Kosti I;Shomron N;Grigoryan V;Przytycka TM;Sauna ZE;Salari R;Mandel-Gutfreund Y;Komar AA;Kimchi-Sarfaty C
通讯作者：
Kimchi-Sarfaty C
DOI：
10.1073/pnas.1304227110
发表时间：
2013-08-13
影响因子：
11.1
作者：
Gartner, Jared J.;Parker, Stephen C. J.;Samuels, Yardena
通讯作者：
Samuels, Yardena
影响因子：
3.9
作者：
Hamasaki-Katagiri, N.;Salari, R.;Kimchi-Sarfaty, C.
通讯作者：
Kimchi-Sarfaty, C.
影响因子：
3.9
作者：
Simhadri, V. L.;Hamasaki-Katagiri, N.;Kimchi-Sarfaty, C.
通讯作者：
Kimchi-Sarfaty, C.
影响因子：
16.8
作者：
Pechmann, Sebastian;Chartron, Justin W.;Frydman, Judith
通讯作者：
Frydman, Judith