Functional analysis of a cardiac myosin rod in Dictyostelium discoideum.

Functional analysis of a cardiac myosin rod in Dictyostelium discoideum.
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盘基网柄菌心肌肌球蛋白棒的功能分析。

DOI:
10.1002/cm.970270404
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发表时间:
1994
影响因子:
--
通讯作者:
Leinwand,LA
Leinwand,LA
中科院分区:
--
文献类型:
--
作者:
LeBlanc-Straceski,JM;Fukui,Y;Sohn,RL;Spudich,JA;Leinwand,LA

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在盘状网柄菌(DD)中对单个常规肌球蛋白重链(MHC)基因的操纵已经描绘出肌球蛋白分子的细丝形成或轻质肌球蛋白(LMM)结构域在细胞运动、发育和细胞表面受体封顶中的重要作用(见Spudich:Cell Regulation 1:1-11,1989;Egelhoff等人:Journal of Cell Biology,112:677-688,1991a)。为了研究肌球蛋白和胞浆肌球蛋白之间的功能关系,将编码肌球蛋白头部和亚片段2的嵌合基因与大鼠β心脏LMM融合,分别导入野生型和DD MHC零细胞。在野生型和DD MHC缺失细胞的细胞质中,嵌合肌球蛋白被组织成致密的皮质斑块。尽管表达嵌合MHC水平为∼10%的空细胞不能在振荡悬浮液中生长或完全发育,但嵌合肌球蛋白能够挽救细胞表面受体的封顶,与丝状肌动蛋白结合,并在聚集过程中定位到正确的亚细胞位置。杆状结构中29个氨基酸的缺失对应于先前定义的细丝组装适配区,消除了趋化过程中的皮质斑块和后部定位。综上所述,这些观察表明,肌节和细胞质肌球蛋白棒在非肌肉运动的几个方面是可以功能互换的。
Manipulation of the single conventional myosin heavy chain (mhc) gene in Dictyostelium discoideum (Dd) has delineated an essential role for the filament-forming, or light meromyosin (LMM) domain of the myosin molecule in cyto-kinesis, development, and in the capping of cell surface receptors (see Spudich: Cell Regulation 1: 1–11, 1989; Egelhoff et al.: Journal of Cell Biology, 112: 677–688, 1991a). In order to assess the functional relationship between sarcomeric and cytoplasmic myosins, a chimeric gene encoding the Dd myosin head and subfragment 2 fused to rat β cardiac LMM was transfected into both wild-type and Dd mhc null cells. Chimeric myosin was organized into dense cortical patches in the cytoplasm of both wild-type and Dd mhc null cells. Although null cells expressing chimeric mhc at∼ 10% of Dd mhc levels were unable to grow in shaking suspension or to complete development, chimeric myosin was able to rescue capping of cell surface receptors, to associate with filamentous actin, and to localize to the correct subcellular position during aggregation. Deletion of 29 amino acids in the rod corresponding to a previously defined filament assembly competent region eliminated the cortical patches and the posterior localization during chemotaxis. Taken together, these observations suggest that sarcomeric and cytoplasmic myosin rods are functionally interchangeable in several aspects of nonmuscle motility.© 1994 Wiley-Liss, Inc.
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