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首页> 外文期刊>Cell motility and the cytoskeleton >Sequence and phylogenetic analysis of squid myosin-V: a vesicle motor in nerve cells.
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Sequence and phylogenetic analysis of squid myosin-V: a vesicle motor in nerve cells.

机译：鱿鱼肌球蛋白-V的序列和系统发育分析：神经细胞中的囊泡运动。

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摘要

We have shown that vesicles in the axoplasm of the squid giant axon move on actin filaments and that movement is inhibited by myosin V-specific antibodies [Tabb et al., 1998]. In the study reported in this article, experiments were performed to clone and sequence the cDNA for squid brain myosin V. Five proteolytic fragments of purified squid brain myosin V were analyzed by direct protein sequencing [Tabb et al., 1998]. Based on this sequence information, degenerate primers were constructed and used to isolate cDNA clones by PCR. Five clones, representing overlapping segments of the gene, were sequenced. The sequence data and the previous biochemical characterization of the molecule support the classification of this vesicle-associated myosin as a member of the class V myosins. Motif analysis of the head, neck, and tail domains revealed that squid MyoV has consensus sequences for all the motifs found in vertebrate members of the myosin V family of motor proteins. A phylogenetic tree was constructed from a sequence alignment by the neighbor-joining method, using Megalign (DNAStar, Madison, WI); the resulting phylogenetic tree showed that squid MyoV is more closely related to vertebrate MyoV (mouse dilute, chicken dilute, rat myr6, and human myo5a) than Drosophila and yeast (myo2, and myo4) myosins V. These new data on the phylogenetic relationships of squid myosin V to vertebrate myosin V strengthens the argument that myosin V functions as a vesicle motor in vertebrate neurons. Copyright 2000 Wiley-Liss, Inc.
机译：我们已经证明，鱿鱼巨型轴突的轴突上的囊泡在肌动蛋白丝上移动，并且该移动受到肌球蛋白V特异性抗体的抑制[Tabb等，1998]。在本文报道的研究中，进行了克隆和测序鱿鱼脑肌球蛋白V cDNA的实验。通过直接蛋白质测序分析了纯化的鱿鱼脑肌球蛋白V的五个蛋白水解片段[Tabb等，1998]。根据该序列信息，构建简并引物并用于通过PCR分离cDNA克隆。对代表该基因重叠片段的五个克隆进行了测序。该分子的序列数据和先前的生化特征支持了该囊泡相关的肌球蛋白分类为V类肌球蛋白的成员。头部，颈部和尾部结构域的基元分析显示，鱿鱼MyoV具有在肌球蛋白V家族运动蛋白家族脊椎动物成员中发现的所有基序的共有序列。使用Megalign（DNAStar，Madison，WI），通过邻居结合法由序列比对构建系统发育树。所形成的系统发育树显示，鱿鱼MyoV与脊椎动物MyoV（小鼠稀稀，鸡肉稀稀，大鼠myr6和人类myo5a）的关系比果蝇和酵母（myo2和myo4）的肌球蛋白V更紧密。这些新数据涉及鱿鱼肌球蛋白V与脊椎动物肌球蛋白V的关系更强，认为肌球蛋白V在脊椎动物神经元中起着囊泡运动的作用。版权所有2000 Wiley-Liss，Inc.

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