Gangwani Laxman, Flavell Richard A, Davis Roger J
Program in Molecular Medicine, Howard Hughes Medical Institute, University of Massachusetts Medical School, 373 Plantation St., Worcester, MA 01605, USA.
Mol Cell Biol. 2005 Apr;25(7):2744-56. doi: 10.1128/MCB.25.7.2744-2756.2005.
Mutation of the survival motor neurons 1 (SMN1) gene causes motor neuron apoptosis and represents the major cause of spinal muscular atrophy in humans. Biochemical studies have established that the SMN protein plays an important role in spliceosomal small nuclear ribonucleoprotein (snRNP) biogenesis and that the SMN complex can interact with the zinc finger protein ZPR1. Here we report that targeted ablation of the Zpr1 gene in mice disrupts the subcellular localization of both SMN and spliceosomal snRNPs. Specifically, SMN localization to Cajal bodies and gems was not observed in cells derived from Zpr1-/- embryos and the amount of cytoplasmic snRNP detected in Zpr1-/- embryos was reduced compared with that in wild-type embryos. We found that Zpr1-/- mice die during early embryonic development, with reduced proliferation and increased apoptosis. These effects of Zpr1 gene disruption were confirmed and extended in studies of cultured motor neuron-like cells using small interfering RNA-mediated Zpr1 gene suppression; ZPR1 deficiency caused growth cone retraction, axonal defects, and apoptosis. Together, these data indicate that ZPR1 contributes to the regulation of SMN complexes and that it is essential for cell survival.
生存运动神经元1(SMN1)基因突变会导致运动神经元凋亡,是人类脊髓性肌萎缩症的主要病因。生化研究表明,SMN蛋白在剪接体小核核糖核蛋白(snRNP)生物合成中起重要作用,且SMN复合物能与锌指蛋白ZPR1相互作用。在此,我们报道在小鼠中靶向敲除Zpr1基因会破坏SMN和剪接体snRNP的亚细胞定位。具体而言,在来自Zpr1-/-胚胎的细胞中未观察到SMN定位于 Cajal体和宝石样小体,且与野生型胚胎相比,在Zpr1-/-胚胎中检测到的细胞质snRNP量减少。我们发现Zpr1-/-小鼠在胚胎发育早期死亡,增殖减少且凋亡增加。利用小干扰RNA介导的Zpr1基因抑制对培养的运动神经元样细胞进行研究,证实并扩展了Zpr1基因破坏的这些效应;ZPR1缺乏导致生长锥回缩、轴突缺陷和凋亡。这些数据共同表明,ZPR1有助于调节SMN复合物,且对细胞存活至关重要。