Cargnin Francesca, Nechiporuk Tamilla, Müllendorff Karin, Stumpo Deborah J, Blackshear Perry J, Ballas Nurit, Mandel Gail
Howard Hughes Medical Institute, Vollum Institute, Oregon Health & Science University, Portland, Oregon 97239.
Laboratory of Signal Transduction, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, and.
J Neurosci. 2014 Dec 10;34(50):16650-61. doi: 10.1523/JNEUROSCI.1650-14.2014.
The RE1 Silencing Transcription Factor (REST) acts as a governor of the mature neuronal phenotype by repressing a large consortium of neuronal genes in non-neuronal cells. In the developing nervous system, REST is present in progenitors and downregulated at terminal differentiation to promote acquisition of mature neuronal phenotypes. Paradoxically, REST is still detected in some regions of the adult nervous system, but how REST levels are regulated, and whether REST can still repress neuronal genes, is not known. Here, we report that homeostatic levels of REST are maintained in mature peripheral neurons by a constitutive post-transcriptional mechanism. Specifically, using a three-hybrid genetic screen, we identify the RNA binding protein, ZFP36L2, associated previously only with female fertility and hematopoiesis, and show that it regulates REST mRNA stability. Dorsal root ganglia in Zfp36l2 knock-out mice, or wild-type ganglia expressing ZFP36L2 shRNA, show higher steady-state levels of Rest mRNA and protein, and extend thin and disintegrating axons. This phenotype is due, at least in part, to abnormally elevated REST levels in the ganglia because the axonal phenotype is attenuated by acute knockdown of REST in Zfp36l2 KO DRG explants. The higher REST levels result in lower levels of target genes, indicating that REST can still fine-tune gene expression through repression. Thus, REST levels are titrated in mature peripheral neurons, in part through a ZFP36L2-mediated post-transcriptional mechanism, with consequences for axonal integrity.
RE1沉默转录因子(REST)通过抑制非神经元细胞中大量的神经元基因,充当成熟神经元表型的调控因子。在发育中的神经系统中,REST存在于祖细胞中,并在终末分化时下调,以促进成熟神经元表型的获得。矛盾的是,在成体神经系统的某些区域仍能检测到REST,但REST水平是如何调控的,以及REST是否仍能抑制神经元基因,目前尚不清楚。在此,我们报告成熟外周神经元通过一种组成性的转录后机制维持REST的稳态水平。具体而言,我们利用三杂交遗传筛选,鉴定出了RNA结合蛋白ZFP36L2,该蛋白之前仅与雌性生育力和造血作用相关,并表明它调节REST mRNA的稳定性。Zfp36l2基因敲除小鼠的背根神经节,或表达ZFP36L2 shRNA的野生型神经节,显示出Rest mRNA和蛋白的稳态水平更高,并且轴突细且有崩解现象。这种表型至少部分是由于神经节中REST水平异常升高所致,因为在Zfp36l2基因敲除的背根神经节外植体中,通过急性敲低REST可减弱轴突表型。较高的REST水平导致靶基因水平降低,表明REST仍可通过抑制作用微调基因表达。因此,成熟外周神经元中REST水平受到调控,部分是通过ZFP36L2介导的转录后机制,这对轴突完整性有影响。