Fukushige T, Yasuda H, Siddiqui S S
Department of Ecological Engineering, Toyohashi University of Technology, Japan.
Biochim Biophys Acta. 1995 Apr 26;1261(3):401-16. doi: 10.1016/0167-4781(95)00028-f.
In the nematode Caenorhabditis elegans, a monoclonal antibody 3A5 raised against Drosophila alpha tubulins selectively stains the nervous system immuno-cytochemically. Direct screening of a C. elegans cDNA expression library with 3A5 has allowed cloning of the tba-1 (tubulin alpha-1) gene from C. elegans. The corresponding genomic DNA encodes a protein of 449 amino acid residues that has a high homology with the vertebrate alpha tubulins but a lower homology with yeast alpha tubulins. Interestingly, the carboxyl-terminus sequence EEEGEEY (Glu-Glu-Glu-Gly-Glu-Glu-Tyr) of the nematode tba-1 encoded isotype is identical to these residues in human, mouse, rat, pig and chicken alpha-1 tubulin isotypes that are expressed in the brain. Temporal and spatial expression studies of the tba-1 gene using Northern blot analysis and tba-1::lacZ fusion gene expression analysis during embryonic and the postembryonic development of C. elegans reveal that the tba-1 tubulin is preferentially expressed in the nematode nervous system, especially in a set of mechanosensory neurons and a set of ventral cord motor neurons (DA, DB, VA, and VB) during embryonic and postembryonic development. Our results indicate an inter-species conservation of the alpha tubulin carboxyl-terminal domain in functionally related brain specific isotypes from metazoans as divergent as mammals and nematodes. These results also suggest specificity of the individual alpha tubulin isotypes during neural development.
在秀丽隐杆线虫中,一种针对果蝇α微管蛋白产生的单克隆抗体3A5通过免疫细胞化学方法选择性地标记神经系统。用3A5直接筛选秀丽隐杆线虫cDNA表达文库,使得从秀丽隐杆线虫中克隆出了tba-1(微管蛋白α-1)基因。相应的基因组DNA编码一个含有449个氨基酸残基的蛋白质,该蛋白质与脊椎动物的α微管蛋白具有高度同源性,但与酵母α微管蛋白的同源性较低。有趣的是,线虫tba-1编码的同种型的羧基末端序列EEEGEEY(谷氨酸-谷氨酸-谷氨酸-甘氨酸-谷氨酸-谷氨酸-酪氨酸)与在大脑中表达的人、小鼠、大鼠、猪和鸡的α-1微管蛋白同种型中的这些残基相同。利用Northern印迹分析以及在秀丽隐杆线虫胚胎期和胚后期发育过程中进行的tba-1::lacZ融合基因表达分析,对tba-1基因进行的时空表达研究表明,tba-1微管蛋白在秀丽隐杆线虫神经系统中优先表达,尤其是在胚胎期和胚后期发育过程中的一组机械感觉神经元以及一组腹神经索运动神经元(DA、DB、VA和VB)中。我们的结果表明,后生动物(如哺乳动物和线虫)中功能相关的脑特异性同种型的α微管蛋白羧基末端结构域存在种间保守性。这些结果还表明了单个α微管蛋白同种型在神经发育过程中的特异性。