Zuccato Chiara, Belyaev Nikolai, Conforti Paola, Ooi Lezanne, Tartari Marzia, Papadimou Evangelia, MacDonald Marcy, Fossale Elisa, Zeitlin Scott, Buckley Noel, Cattaneo Elena
Department of Pharmacological Sciences and Centre for Stem Cell Research, University of Milan, Via Balzaretti 9, 20133 Milano, Italy.
J Neurosci. 2007 Jun 27;27(26):6972-83. doi: 10.1523/JNEUROSCI.4278-06.2007.
Huntingtin is a protein that is mutated in Huntington's disease (HD), a dominant inherited neurodegenerative disorder. We previously proposed that, in addition to the gained toxic activity of the mutant protein, selective molecular dysfunctions in HD may represent the consequences of the loss of wild-type protein activity. We first reported that wild-type huntingtin positively affects the transcription of the brain-derived neurotrophic factor (BDNF) gene, a cortically derived survival factor for the striatal neurons that are mainly affected in the disease. Mutation in huntingtin decreases BDNF gene transcription. One mechanism involves the activation of repressor element 1/neuron-restrictive silencer element (RE1/NRSE) located within the BDNF promoter. We now show that increased binding of the RE1 silencing transcription factor/neuron-restrictive silencer factor (REST/NRSF) repressor occurs at multiple genomic RE1/NRSE loci in HD cells, in animal models, and in postmortem brains, resulting in a decrease of RE1/NRSE-mediated gene transcription. The same molecular phenotype is produced in cells and brain tissue depleted of endogenous huntingtin, thereby directly validating the loss-of-function hypothesis of HD. Through a ChIP (chromatin immunoprecipitation)-on-chip approach, we examined occupancy of multiple REST/NRSF target genes in the postmortem HD brain, providing the first example of the application of this technology to neurodegenerative diseases. Finally, we show that attenuation of REST/NRSF binding restores BDNF levels, suggesting that relief of REST/NRSF mediated repression can restore aberrant neuronal gene transcription in HD.
亨廷顿蛋白是一种在亨廷顿舞蹈症(HD)中发生突变的蛋白质,HD是一种显性遗传性神经退行性疾病。我们之前提出,除了突变蛋白获得的毒性活性外,HD中选择性的分子功能障碍可能代表野生型蛋白活性丧失的后果。我们首次报道野生型亨廷顿蛋白对脑源性神经营养因子(BDNF)基因的转录有积极影响,BDNF是一种主要在该疾病中受影响的纹状体神经元的皮质源性存活因子。亨廷顿蛋白的突变会降低BDNF基因的转录。一种机制涉及位于BDNF启动子内的阻遏元件1/神经元限制性沉默元件(RE1/NRSE)的激活。我们现在表明,在HD细胞、动物模型和死后大脑的多个基因组RE1/NRSE位点,RE1沉默转录因子/神经元限制性沉默因子(REST/NRSF)阻遏物的结合增加,导致RE1/NRSE介导的基因转录减少。在内源性亨廷顿蛋白缺失的细胞和脑组织中也产生了相同的分子表型,从而直接验证了HD的功能丧失假说。通过芯片染色质免疫沉淀(ChIP)方法,我们检测了死后HD大脑中多个REST/NRSF靶基因的占据情况,这是该技术应用于神经退行性疾病的首个实例。最后,我们表明REST/NRSF结合的减弱可恢复BDNF水平,这表明解除REST/NRSF介导的抑制可恢复HD中异常的神经元基因转录。