Hervé D, Rogard M, Lévi-Strauss M
INSERM U114, Chaire de Neuropharmacologie, Collège de France, Paris, France.
Brain Res Mol Brain Res. 1995 Aug;32(1):125-34. doi: 10.1016/0169-328x(95)00070-9.
The alpha subunit of GTP-binding protein Golf (G alpha olf), identified in the olfactory epithelium, in which it is supposed to mediate odorant-generated adenylyl cyclase activations, is much more expressed in the striatum than G alpha s, the classical stimulatory G protein alpha subunit (Hervé et al., J Neurosci., 13 (1993) 2237-2248). Four species of G alpha olf messenger RNA (mRNA) were observed by Northern blot in the rat striatum. Analysis of striatal G alpha olf cDNA clones demonstrated that despite extensive variations in their 5' and 3' untranslated regions, these four G alpha olf mRNAs encode the same G alpha olf polypeptide. One of the four mRNA species, which was selectively observed in the striatum, is generated by a novel promoter whose activity was barely detectable in the olfactory epithelium. Surprisingly, this promoter generates an unexpectedly high proportion of transcripts in which the first intron is unspliced. The retention of intron was found to abolish the translation of G alpha olf mRNA in the reticulocyte lysate system, suggesting that it could be involved in the down regulation of G alpha olf expression in the striatum. Furthermore, a new polyadenylation site with a non canonical sequence, AATACA, was found to be responsible of the two shortest forms of G alpha olf mRNA. In conclusion, we have shown that the G alpha olf proteins present in the striatum and the olfactory epithelium are identical and that multiple variations in the untranslated parts of the mRNAs could affect G alpha olf expression.
在嗅觉上皮中发现的GTP结合蛋白Golf的α亚基(Gαolf),被认为在其中介导气味剂诱导的腺苷酸环化酶激活,它在纹状体中的表达远高于经典的刺激性G蛋白α亚基Gαs(埃尔韦等人,《神经科学杂志》,13卷(1993年)2237 - 2248页)。通过Northern印迹法在大鼠纹状体中观察到四种Gαolf信使核糖核酸(mRNA)。对纹状体Gαolf互补脱氧核糖核酸(cDNA)克隆的分析表明,尽管它们的5'和3'非翻译区存在广泛差异,但这四种Gαolf mRNA编码相同的Gαolf多肽。在纹状体中选择性观察到的四种mRNA之一,是由一个新的启动子产生的,该启动子的活性在嗅觉上皮中几乎检测不到。令人惊讶的是,这个启动子产生的转录本中,第一个内含子未剪接的比例出乎意料地高。发现内含子的保留会消除Gαolf mRNA在网织红细胞裂解物系统中的翻译,这表明它可能参与纹状体中Gαolf表达的下调。此外,发现一个具有非规范序列AATACA的新聚腺苷酸化位点,负责产生两种最短形式的Gαolf mRNA。总之,我们已经表明,纹状体和嗅觉上皮中存在的Gαolf蛋白是相同的,并且mRNA非翻译部分的多种变化可能影响Gαolf的表达。