Zhang Meiqin, Wang Qiaoqiao, Huang Yingqun
Department of Obstetrics, Gynecology, and Reproductive Sciences, Yale University School of Medicine, 300 George Street, New Haven, CT 06511, USA.
Proc Natl Acad Sci U S A. 2007 Jun 12;104(24):10057-62. doi: 10.1073/pnas.0700169104. Epub 2007 Jun 4.
Fragile X syndrome is caused by the inactivation of the X-linked FMR1 gene, leading to the loss of its encoded protein FMRP. Although macroorchidism and defects in neuronal architecture and function have been associated with lack of FMRP, the exact molecular mechanism underlying this disease remains unclear. We have reported previously that in the brain and testis of mice, FMRP specifically interacts with a distinct mRNA nuclear export factor NXF2 but not with its close relative NXF1, a ubiquitously expressed essential mRNA nuclear export factor. This interaction marked NXF2 as a putative functional partner of FMRP. Here, we demonstrate by immunoprecipitation and quantitative real-time RT-PCR that, in cultured mouse neuronal cells, both FMRP and NXF2 are present in Nxf1 mRNA-containing ribonucleoprotein particles. Further, we show that expression of NXF2 leads to the destabilization of Nxf1 mRNA and that this effect is abolished when Fmr1 expression is reduced by siRNA, arguing that both proteins collaborate to exert this effect. Importantly, these findings correlate well with our observations that in both mouse hippocampal neurons and male germ cells where the expression of FMRP and NXF2 is most prominent, the expression of NXF1 is relatively poorly expressed. Our studies thus identify Nxf1 mRNA as a likely biologically relevant in vivo target of both FMRP and NXF2 and implicate FMRP, in conjunction with NXF2, as a posttranscriptional regulator of a major mRNA export factor. Such regulation may prove important in the normal development and function of neurons as well as of male germ cells.
脆性X综合征是由X连锁的FMR1基因失活引起的,导致其编码的蛋白质FMRP缺失。尽管巨睾症以及神经元结构和功能缺陷与FMRP的缺乏有关,但这种疾病的确切分子机制仍不清楚。我们之前报道过,在小鼠的大脑和睾丸中,FMRP特异性地与一种独特的mRNA核输出因子NXF2相互作用,而不与其关系密切的NXF1相互作用,NXF1是一种普遍表达的必需mRNA核输出因子。这种相互作用将NXF2标记为FMRP的假定功能伙伴。在此,我们通过免疫沉淀和定量实时RT-PCR证明,在培养的小鼠神经元细胞中,FMRP和NXF2都存在于含有Nxf1 mRNA的核糖核蛋白颗粒中。此外,我们表明NXF2的表达导致Nxf1 mRNA的不稳定,并且当通过siRNA降低Fmr1表达时这种效应被消除,这表明这两种蛋白质共同发挥这种作用。重要的是,这些发现与我们的观察结果高度相关,即在FMRP和NXF2表达最突出的小鼠海马神经元和雄性生殖细胞中,NXF1的表达相对较低。因此,我们的研究确定Nxf1 mRNA可能是FMRP和NXF2在体内的生物学相关靶点,并表明FMRP与NXF2一起作为主要mRNA输出因子的转录后调节因子。这种调节可能在神经元以及雄性生殖细胞的正常发育和功能中被证明是重要的。