Linial M, Scheller R H
Department of Biological Sciences, Stanford University, CA 94305.
J Neurochem. 1990 Mar;54(3):762-70. doi: 10.1111/j.1471-4159.1990.tb02316.x.
A set of cDNA clones encoding a protein highly homologous to the mammalian middle-size class of neurofilaments (NF-M) was characterized. The amino acid similarity between the Torpedo and rat NF-M approaches 90% in the amino-terminal "rod-like" domain and is significantly lower in the carboxy-terminal tail. The Torpedo protein contains 13 tandem repeats of a unique six amino acid core, containing a Lys-Ser-Lys putative phosphorylation site. Surprisingly, the 3' untranslated region contains stretches of 80-90% nucleic acid homology with the mammalian, but not with the chicken sequences. This homology is greater than much of the coding region, suggesting that the 3' untranslated region of the message has an important functional role, perhaps governing RNA stability or localization. This Torpedo NF-M mRNA is expressed specifically in the electric lobe and was not detected in other tissues, including brain and spinal cord. A polyclonal antibody generated against a fusion protein synthesized in E. coli detects a 150-kDa protein in the electric lobe and organ, as well as a small amount of material in the brain. Cytochemical studies reveal immunoreactivity in electromotor neuron axons and terminals. Specific expression of neurofilament genes in subsets of central neurons may be important in determining the morphology and functional characteristics of specific neuronal subtypes.
对一组编码与哺乳动物中等大小神经丝(NF-M)高度同源蛋白质的cDNA克隆进行了表征。电鳐和大鼠NF-M在氨基末端“杆状”结构域的氨基酸相似性接近90%,而在羧基末端尾部则显著降低。电鳐蛋白含有13个独特的六氨基酸核心串联重复序列,其中包含一个赖氨酸-丝氨酸-赖氨酸假定磷酸化位点。令人惊讶的是,3'非翻译区与哺乳动物序列有80-90%的核酸同源性片段,但与鸡的序列没有。这种同源性大于许多编码区,表明该信使RNA的3'非翻译区具有重要的功能作用,可能控制RNA的稳定性或定位。这种电鳐NF-M mRNA在电叶中特异性表达,在包括脑和脊髓在内的其他组织中未检测到。针对在大肠杆菌中合成的融合蛋白产生的多克隆抗体,在电叶和器官中检测到一种150 kDa的蛋白质,在脑中也检测到少量物质。细胞化学研究揭示了在运动神经元轴突和终末的免疫反应性。神经丝基因在中枢神经元亚群中的特异性表达可能对确定特定神经元亚型的形态和功能特征很重要。