Endo T, Matsumoto K, Hama T, Ohtsuka Y, Katsura G, Obinata T
Department of Biology, Faculty of Science, Chiba University, Yayoicho, Inageku, Chiba 263, Japan.
J Biol Chem. 1996 Nov 1;271(44):27855-62. doi: 10.1074/jbc.271.44.27855.
During development of the ascidian Halocynthia roretzi, the tadpole larva hatched from the tailbud embryo metamorphoses to the sessile adult with a body wall muscle. Although the adult body wall muscle is morphologically nonsarcomeric smooth muscle, it contains troponin complex consisting of three subunits (T, I, and C) as do vertebrate striated muscles. Different from vertebrate troponins, however, the smooth muscle troponin promotes actomyosin Mg2+-ATPase activity in the presence of high concentration of Ca2+, and this promoting property is attributable to troponin T. To address whether the embryonic/larval tail striated muscle and the adult smooth muscle utilize identical or different regulatory machinery, we cloned troponin T cDNAs from each cDNA library. The embryonic and the adult troponin Ts were encoded by distinct genes and shared only <60% identity with each other. Northern blotting and whole mount in situ hybridization revealed that these isoforms were specifically expressed in the embryonic/larval tail striated muscle and the adult smooth muscle, respectively. These results may imply that these isoforms regulate actin-myosin interaction in different manners. The adult troponin T under forced expression in mouse fibroblasts was unexpectedly located in the nuclei. However, a truncated protein with a deletion including a cluster of basic amino acids colocalized with tropomyosin on actin filaments. Thus, complex formation with troponin I and C immediately after the synthesis is likely to be essential for the protein to properly localize on the thin filaments.
在海鞘柄海鞘发育过程中,从尾芽胚胎孵化出的蝌蚪幼虫会变态发育为具有体壁肌肉的固着成体。虽然成体的体壁肌肉在形态上是非肌节性平滑肌,但它像脊椎动物的横纹肌一样,含有由三个亚基(T、I和C)组成的肌钙蛋白复合体。然而,与脊椎动物的肌钙蛋白不同的是,平滑肌肌钙蛋白在高浓度Ca2+存在的情况下会促进肌动球蛋白Mg2+-ATP酶的活性,而且这种促进特性归因于肌钙蛋白T。为了探究胚胎/幼虫尾部的横纹肌与成体的平滑肌是否利用相同或不同的调节机制,我们从每个cDNA文库中克隆了肌钙蛋白T的cDNA。胚胎型和成体型肌钙蛋白T由不同的基因编码,彼此之间的同源性仅<60%。Northern印迹法和整体原位杂交显示,这些同工型分别在胚胎/幼虫尾部的横纹肌和成体的平滑肌中特异性表达。这些结果可能意味着这些同工型以不同的方式调节肌动蛋白-肌球蛋白的相互作用。意外的是,在小鼠成纤维细胞中强制表达的成体肌钙蛋白T定位于细胞核中。然而,一种缺失了包括一簇碱性氨基酸的截短蛋白与原肌球蛋白在肌动蛋白丝上共定位。因此,合成后立即与肌钙蛋白I和C形成复合体可能是该蛋白正确定位于细肌丝上所必需的。