From the Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260.
J Biol Chem. 2014 May 2;289(18):12727-47. doi: 10.1074/jbc.M113.529123. Epub 2014 Mar 13.
Early-onset torsion dystonia (EOTD) is a neurological disorder characterized by involuntary and sustained muscle contractions that can lead to paralysis and abnormal posture. EOTD is associated with the deletion of a glutamate (ΔE) in torsinA, an endoplasmic reticulum (ER) resident AAA(+) ATPase. To date, the effect of ΔE on torsinA and the reason that this mutation results in EOTD are unclear. Moreover, there are no specific therapeutic options to treat EOTD. To define the underlying biochemical defects associated with torsinAΔE and to uncover factors that might be targeted to offset defects associated with torsinAΔE, we developed a yeast torsinA expression system and tested the roles of ER chaperones in mediating the folding and stability of torsinA and torsinAΔE. We discovered that the ER lumenal Hsp70, BiP, an associated Hsp40, Scj1, and a nucleotide exchange factor, Lhs1, stabilize torsinA and torsinAΔE. BiP also maintained torsinA and torsinAΔE solubility. Mutations predicted to compromise specific torsinA functional motifs showed a synthetic interaction with the ΔE mutation and destabilized torsinAΔE, suggesting that the ΔE mutation predisposes torsinA to defects in the presence of secondary insults. In this case, BiP was required for torsinAΔE degradation, consistent with data that specific chaperones exhibit either pro-degradative or pro-folding activities. Finally, using two independent approaches, we established that BiP stabilizes torsinA and torsinAΔE in mammalian cells. Together, these data define BiP as the first identified torsinA chaperone, and treatments that modulate BiP might improve symptoms associated with EOTD.
早发性扭转痉挛(EOTD)是一种以不自主和持续肌肉收缩为特征的神经障碍,可导致瘫痪和异常姿势。EOTD 与 torsinA 中谷氨酸(ΔE)的缺失有关,torsinA 是内质网(ER)驻留的 AAA(+)ATP 酶。迄今为止,ΔE 对 torsinA 的影响以及这种突变导致 EOTD 的原因尚不清楚。此外,目前尚无治疗 EOTD 的特定方法。为了确定与 torsinAΔE 相关的潜在生化缺陷,并揭示可能用于抵消与 torsinAΔE 相关缺陷的因素,我们开发了一种酵母 torsinA 表达系统,并测试了 ER 伴侣在介导 torsinA 和 torsinAΔE 折叠和稳定性方面的作用。我们发现 ER 腔中的 Hsp70、伴侣蛋白 BiP、相关的 Hsp40 Scj1 和核苷酸交换因子 Lhs1 稳定 torsinA 和 torsinAΔE。BiP 还维持 torsinA 和 torsinAΔE 的可溶性。预测会损害特定 torsinA 功能基序的突变与 ΔE 突变表现出合成相互作用,并使 torsinAΔE 不稳定,这表明在存在二次损伤的情况下,ΔE 突变使 torsinA 易于出现缺陷。在这种情况下,BiP 是 torsinAΔE 降解所必需的,这与特定伴侣蛋白表现出促降解或促折叠活性的数据一致。最后,我们使用两种独立的方法证实 BiP 在哺乳动物细胞中稳定 torsinA 和 torsinAΔE。这些数据共同将 BiP 定义为第一个鉴定出的 torsinA 伴侣蛋白,调节 BiP 的治疗方法可能会改善与 EOTD 相关的症状。