Buiakova O I, Krishna N S, Getchell T V, Margolis F L
Roche Institute of Molecular Biology, Nutley, New Jersey 07110.
Genomics. 1994 Apr;20(3):452-62. doi: 10.1006/geno.1994.1200.
Immunocytochemical analysis has demonstrated that expression of the olfactory marker protein (OMP) is highly restricted to mature olfactory receptor neurons in virtually all vertebrate species from fish to man. We have now cloned the OMP gene from human and mouse and demonstrated conservation of gene structure, protein sequence, and Olf-1 and upstream binding region (UBE) regulatory domains. The OMP gene in all species studied lacks canonical TATA and CAAT motifs and introns. The deduced protein sequence is 88.4% identical between mouse and human, and most of the differences observed are conservative changes. The proximal Olf-1 binding sites differ by two purine-purine replacements and effectively cross-compete in mobility shift assays. The distal Olf-1 binding site is also highly conserved in terms of both sequence and binding activity. The availability of sequence from multiple species has permitted us to determine that the UBE site has close similarity to motifs that bind members of the NF-1 family of transcription factors. Gel mobility shift assays confirm this prediction, providing additional insight into mechanisms that may participate in the stringent regulation of the expression of this neuronal-specific protein. Furthermore, we demonstrate the in situ localization of OMP mRNA in human olfactory neuro-epithelium and its colocalization to immunocytochemically identified human olfactory receptor neurons.
免疫细胞化学分析表明,嗅觉标记蛋白(OMP)的表达在从鱼类到人类的几乎所有脊椎动物物种中都高度局限于成熟的嗅觉受体神经元。我们现已从人和小鼠中克隆出OMP基因,并证明了基因结构、蛋白质序列以及Olf-1和上游结合区域(UBE)调控域的保守性。在所研究的所有物种中,OMP基因均缺乏典型的TATA和CAAT基序以及内含子。推导的小鼠和人类蛋白质序列的同源性为88.4%,观察到的大多数差异都是保守性变化。近端Olf-1结合位点有两个嘌呤-嘌呤置换差异,并且在迁移率变动分析中能有效交叉竞争。远端Olf-1结合位点在序列和结合活性方面也高度保守。多个物种的序列信息使我们能够确定UBE位点与结合转录因子NF-1家族成员的基序非常相似。凝胶迁移率变动分析证实了这一预测,为可能参与这种神经元特异性蛋白表达严格调控的机制提供了更多见解。此外,我们证明了OMP mRNA在人嗅觉神经上皮中的原位定位及其与免疫细胞化学鉴定的人嗅觉受体神经元的共定位。