Sherr E H, Joyce M P, Greene L A
Department of Pathology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
J Cell Biol. 1993 Mar;120(6):1405-16. doi: 10.1083/jcb.120.6.1405.
A polymerase chain reaction strategy was devised to identify new members of the mammalian myosin I family of actin-based motors. Using cellular RNA from mouse granular neurons and PC12 cells, we have cloned and sequenced three 1.2-kb polymerase chain reaction products that correspond to novel mammalian myosin I genes designated MMI alpha, MMI beta, MMI gamma. The pattern of expression for each of the myosin I's is unique: messages are detected in diverse tissues including the brain, lung, kidney, liver, intestine, and adrenal gland. Overlapping clones representing full-length cDNAs for MMI alpha were obtained from mouse brain. These encode a 1,079 amino acid protein containing a myosin head, a domain with five calmodulin binding sites, and a positively charged COOH-terminal tail. In situ hybridization reveals that MMI alpha is highly expressed in virtually all neurons (but not glia) in the postnatal and adult mouse brain and in neuroblasts of the cerebellar external granular layer. Expression varies in different brain regions and undergoes developmental regulation. Myosin I's are present in diverse organisms from protozoa to vertebrates. This and the expression of three novel members of this family in brain and other mammalian tissues suggests that they may participate in critical and fundamental cellular processes.
设计了一种聚合酶链反应策略来鉴定基于肌动蛋白的哺乳动物肌球蛋白I家族的新成员。利用来自小鼠颗粒神经元和PC12细胞的细胞RNA,我们克隆并测序了三个1.2kb的聚合酶链反应产物,它们对应于新的哺乳动物肌球蛋白I基因,命名为MMIα、MMIβ、MMIγ。每种肌球蛋白I的表达模式都是独特的:在包括脑、肺、肾、肝、肠和肾上腺在内的多种组织中都能检测到其信使RNA。从小鼠脑中获得了代表MMIα全长cDNA的重叠克隆。这些克隆编码一种1079个氨基酸的蛋白质,该蛋白质含有一个肌球蛋白头部、一个具有五个钙调蛋白结合位点的结构域和一个带正电荷的COOH末端尾巴。原位杂交显示,MMIα在出生后和成年小鼠脑中的几乎所有神经元(而非神经胶质细胞)以及小脑外颗粒层的神经母细胞中高度表达。其在不同脑区的表达有所不同,并受到发育调控。肌球蛋白I存在于从原生动物到脊椎动物的多种生物体中。该家族三个新成员在脑和其他哺乳动物组织中的这种表达情况表明,它们可能参与关键且基本的细胞过程。