Singh Kanchan, Prasad S
Biochemistry & Molecular Biology laboratory, CAS in Zoology, Banaras Hindu University, Varanasi, 221005, India.
Mol Biol Rep. 2008 Dec;35(4):677-84. doi: 10.1007/s11033-007-9140-0. Epub 2007 Sep 27.
Fragile X syndrome is caused by silencing of FMR-1 gene due to unusual expansion of CGG repeats (>200 repeats) and their hypermethylation in 5'-UTR. As a consequence, the expression of the RNA binding protein FMRP is stopped. Absence of this protein leads to several morphological and neurological symptoms. The symptoms of the syndrome in males are different than that in the females. We have previously reported that the Fmr1 gene is down regulated in males as a function of age. In the present communication, we have investigated expression of Fmr-1 mRNA, FMRP and analysis of interaction of trans-acting factors with E- and GC boxes in Fmr-1 promoter in female mouse brain as a function of age. Our Northern and Western blots data reveal that the level of Fmr-1 transcript decreases in adult as compared to young mouse but significantly increases in old age and that of FMRP decreases in brain of female old mouse as compared to young and adult age. The immunohistochemical analysis supported the results obtained from Western blot studies. Our EMSA data reveal that the intensity of USF1/USF2-E Box complex gradually increases during aging having significantly highest intensity in old age mouse whereas the intensity of alpha-Pal/Nrf1- GC-Box complex gradually decreases as a function of age. The increased intensity of the complex in old age mouse is correlated to higher level of Fmr-1 transcript in old age. The elevated level of Fmr-1 transcript in old mouse brain may be attributed to USF1/USF2 dependent increased transcription of Fmr-1 gene in old age and decrease in FMRP to altered translation of the transcript or high turn over of FMRP during aging. The present finding indicates age and sex as factors affecting the expression of Fmr-1 gene in mouse brain.
脆性X综合征是由于5'-UTR区域中CGG重复序列异常扩增(>200次重复)及其高甲基化导致FMR-1基因沉默所致。因此,RNA结合蛋白FMRP的表达停止。该蛋白的缺失会导致多种形态学和神经学症状。该综合征在男性和女性中的症状有所不同。我们之前报道过,雄性小鼠中Fmr1基因的表达随年龄增长而下调。在本报告中,我们研究了雌性小鼠大脑中Fmr-1 mRNA、FMRP的表达以及Fmr-1启动子中反式作用因子与E盒和GC盒相互作用随年龄的变化情况。我们的Northern印迹和Western印迹数据显示,与幼年小鼠相比,成年小鼠中Fmr-1转录本水平降低,但在老年时显著增加,而老年雌性小鼠大脑中FMRP的水平与幼年和成年时相比降低。免疫组织化学分析支持了Western印迹研究的结果。我们的电泳迁移率变动分析(EMSA)数据显示,USF1/USF2-E盒复合物的强度在衰老过程中逐渐增加,在老年小鼠中强度最高;而α-Pal/Nrf1-GC盒复合物的强度随年龄增长逐渐降低。老年小鼠中复合物强度的增加与老年时Fmr-1转录本水平的升高相关。老年小鼠大脑中Fmr-1转录本水平的升高可能归因于老年时USF1/USF2依赖性Fmr-1基因转录增加,以及FMRP减少导致转录本翻译改变或衰老过程中FMRP的高周转率。目前的研究结果表明年龄和性别是影响小鼠大脑中Fmr-1基因表达的因素。