Laboratory of Molecular Genetics, G. Gaslini Institute, 16148, Genoa, Italy.
J Mol Med (Berl). 2012 Sep;90(9):1025-35. doi: 10.1007/s00109-012-0868-1. Epub 2012 Feb 4.
Expansions of a polyalanine (polyA) stretch in the coding region of the PHOX2B gene cause congenital central hypoventilation syndrome (CCHS), a neurocristopathy characterized by the absence of adequate control of autonomic breathing. Expansion of polyA in PHOX2B leads to protein misfolding and accumulation into inclusions. The mechanisms that regulate mutant protein degradation and turnover have been poorly elucidated. Here, we investigate the regulation of degradation of wild-type and polyA-expanded PHOX2B. We show that expanded PHOX2B is targeted for degradation through the ubiquitin-proteasome system, resulting in lowered levels of the mutant protein relative to its wild-type counterpart. Moreover, we show that mutant PHOX2B forms ubiquitin-positive inclusions, which sequester wild-type PHOX2B. This sequestration correlates with reduced transcriptional activity of endogenous wild-type protein in neuroblastoma cells. Finally, we show that the E3 ubiquitin ligase TRIM11 plays a critical role in the clearance of mutant PHOX2B through the proteasome. Importantly, clearance of mutant PHOX2B by TRIM11 correlates with a rescue of PHOX2B transcriptional activity. We propose that CCHS is partially caused by a dominant-negative effect of expanded PHOX2B due to the retention of the wild-type protein in pathogenic aggregates. Our results demonstrate that TRIM11 is a novel modifier of mutant PHOX2B toxicity and represents a potential therapeutic target for CCHS.
PHOX2B 基因编码区的多聚丙氨酸(polyA)扩展导致先天性中枢性低通气综合征(CCHS),这是一种神经嵴病,其特征是自主呼吸控制不足。PHOX2B 中的 polyA 扩展导致蛋白质错误折叠和积累成包含物。调节突变蛋白降解和周转的机制尚未得到充分阐明。在这里,我们研究了野生型和 polyA 扩展的 PHOX2B 降解的调节。我们表明,扩展的 PHOX2B 通过泛素-蛋白酶体系统被靶向降解,导致突变蛋白的水平相对于其野生型对应物降低。此外,我们表明突变 PHOX2B 形成泛素阳性包含物,这些包含物将野生型 PHOX2B 隔离。这种隔离与神经母细胞瘤细胞中内源性野生型蛋白转录活性的降低相关。最后,我们表明 E3 泛素连接酶 TRIM11 通过蛋白酶体在清除突变 PHOX2B 中发挥关键作用。重要的是,TRIM11 对突变 PHOX2B 的清除与 PHOX2B 转录活性的恢复相关。我们提出,CCHS 部分是由于致病性聚集体中野生型蛋白的保留导致扩展的 PHOX2B 产生的显性负效应所致。我们的结果表明,TRIM11 是突变 PHOX2B 毒性的新型调节剂,代表了 CCHS 的潜在治疗靶点。