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对腹足纲动物欧洲大蜗牛神经丝蛋白NF70 mRNA的表征显示,脑神经节的神经元和非神经元中间丝蛋白源自不同的层粘连相关基因。

Characterisation of neurofilament protein NF70 mRNA from the gastropod Helix aspersa reveals that neuronal and non-neuronal intermediate filament proteins of cerebral ganglia arise from separate lamin-related genes.

作者信息

Adjaye J, Plessmann U, Weber K, Dodemont H

机构信息

Max Planck Institute for Biophysical Chemistry, Department of Biochemistry, Goettingen, Germany.

出版信息

J Cell Sci. 1995 Nov;108 ( Pt 11):3581-90. doi: 10.1242/jcs.108.11.3581.

Abstract

The neuronal cytoplasmic intermediate filament (IF) protein HeNF70 of the gastropod Helix aspersa is identified by sequence analysis of the corresponding 4,600 bp cDNA isolated from a cerebral ganglion cDNA library. HeNF70 shares 60% sequence identity with the neuronal LoNF70 protein of the cephalopod Loligo pealei and only 36% identity with the Helix non-neuronal IF-A protein. All three molluscan IF proteins display the lamin-type extended coil 1b subdomain harbouring six additional heptads and all have long C-terminal sequences with substantial homology to the tail domains of nuclear lamins. The lamin-like tail domains of the two neurofilament proteins share a unique motif comprising 13 residues, which is absent from Helix IF-A and all other known non-neuronal IF proteins. HeNF70 is encoded by a 9.5 kb RNA transcript. The very long 7.2 kb 3'-untranslated sequence contains a unique 26 nucleotide repeat extending over 0.5 kb in its 5'-region. The HeNF70 mRNA is expressed at low abundancy in cerebral ganglia but not in any of the 13 non-neuronal tissues tested. In contrast, all tissues express at fairly high levels the same 4.6 and 4.2 kb mRNAs encoding the Helix non-neuronal IF-A/B proteins. Blot hybridisation studies on genomic DNA and ganglion mRNA with subprobes from the cloned HeNF70 cDNA, as well as sequence analysis of an RT-PCR generated partial cDNA encoding a putative HeNF60 protein, indicate at least two different neuronal IF genes in Helix.

摘要

通过对从脑神经节cDNA文库中分离出的相应4600bp cDNA进行序列分析,鉴定出腹足纲动物皱疤坚螺的神经元细胞质中间丝(IF)蛋白HeNF70。HeNF70与头足纲动物皮氏枪乌贼的神经元LoNF70蛋白有60%的序列同一性,而与皱疤坚螺的非神经元IF-A蛋白只有36%的同一性。所有这三种软体动物IF蛋白都显示出具有六个额外七肽的核纤层蛋白型延伸卷曲1b亚结构域,并且都具有与核纤层蛋白尾结构域有大量同源性的长C末端序列。两种神经丝蛋白的核纤层蛋白样尾结构域共享一个由13个残基组成的独特基序,皱疤坚螺IF-A和所有其他已知的非神经元IF蛋白中都没有这个基序。HeNF70由一个9.5kb的RNA转录本编码。非常长的7.2kb 3'非翻译序列在其5'区域包含一个独特的26个核苷酸的重复序列,延伸超过0.5kb。HeNF70 mRNA在脑神经节中低丰度表达,但在所测试的13个非神经元组织中均未表达。相反,所有组织都以相当高的水平表达相同的4.6kb和4.2kb mRNA,它们编码皱疤坚螺的非神经元IF-A/B蛋白。用克隆的HeNF70 cDNA的亚探针进行的基因组DNA和神经节mRNA的印迹杂交研究,以及对一个推定的HeNF60蛋白的RT-PCR产生的部分cDNA的序列分析,表明皱疤坚螺中至少有两个不同的神经元IF基因。

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