Involvement of the 3′-untranslated region of the brain-derived neurotrophic factor gene in activity-dependent mRNA stabilization
Involvement of the 3′-untranslated region of the brain-derived neurotrophic factor gene in activity-dependent mRNA stabilization
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DOI：
10.1111/j.1471-4159.2010.07016.x
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发表时间：
2010-12-01
影响因子：
4.7
通讯作者：
Tsuda, Masaaki
中科院分区：
文献类型：
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作者：
Fukuchi, Mamoru;Tsuda, Masaaki
虽然基因转录受神经元活动的控制，但对神经元中的转录后调控知之甚少。使用培养的神经元，我们发现，立即早期基因转录本的半衰期延长或缩短膜去极化。为了研究脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）mRNA的活性依赖性稳定性，我们构建了一系列将BDNF基因的3 '-非翻译区（3'-untranslated region，3 '-UTR）与萤火虫荧光素酶基因融合的质粒，发现3'-UTR通过去极化引起的Ca 2+信号阻止荧光素酶mRNA的不稳定性。在猿猴病毒40晚期poly（A）位点没有观察到这种预防作用。覆盖整个短3 '-UTR的前mRNA被稳定化，其中多个poly（A）位点（包括新的位点）位于该短3'-UTR。3 '-UTR的缺失分析揭示了一个核心区（约130个碱基长）和一个互补区负责预防，与延伸的茎环RNA结构的形成和poly（A）mRNA的产生完全一致。因此，mRNA的稳定性在神经元中是活性依赖性控制的，并且BDNF mRNA的3 '-UTR的不同区域参与稳定mRNA以响应Ca 2+信号，这表明RNA二级结构在活性依赖性mRNA稳定中影响多聚腺苷酸位点的可用性的主要作用。
Although gene transcription is controlled by neuronal activity, little is known about post-transcriptional regulation in neurons. Using cultured neurons, we found that the half-life of immediate-early gene transcripts is prolonged or shortened by membrane depolarization. Focusing on the activity-dependent stabilization of brain-derived neurotrophic factor (BDNF) mRNA, we constructed a series of plasmids, in which the short 3'-untranslated region (3'-UTR) of the BDNF gene was fused to the firefly luciferase gene, and found that the 3'-UTR prevented destabilization of luciferase mRNA through Ca2+ signals evoked via depolarization. No such prevention was observed with the simian virus 40 late poly(A) site. The pre-mRNA covering the entire short 3'-UTR, where multiple poly(A) sites including novel ones are located, was stabilized. Deletion analyses of 3'-UTR revealed a core region (about 130 bases long) and a complementary region to be responsible for the prevention, well consistent with the formation of an extended stem-loop RNA structure and the production of poly(A) mRNAs. Thus, the mRNA stability is activity-dependently controlled in neurons and distinct regions of the 3'-UTR of BDNF mRNA are involved in stabilizing mRNA in response to Ca2+ signals, suggesting a primary role of the RNA secondary structure affecting the availability of poly(A) sites in activity-dependent mRNA stabilization.