Antisense suppression of skeletal muscle myosin light chain-1 biosynthesis impairs myofibrillogenesis in cultured myotubes.
Antisense suppression of skeletal muscle myosin light chain-1 biosynthesis impairs myofibrillogenesis in cultured myotubes.
复制标题
骨骼肌肌球蛋白轻链 1 生物合成的反义抑制会损害培养的肌管中的肌原纤维生成。
DOI：
10.1007/bf00125309
复制
发表时间：
1995
影响因子：
2.7
通讯作者：
Mikawa,T
中科院分区：
文献类型：
--
作者：
Nawrotzki,R;Fischman,DA;Mikawa,T
Although the alkali or essential light chains of skeletal muscle myosin are not required for actin-activated myosin ATPase activity, these myosin subunits are necessary for force transmission within vitroactin motility assays and are believed to stabilize the α-helical neck region of myosin subfragment-1. To probe the functions of the essential light chains during myofibril assembly, we used recombinant DNA procedures to deplete this light chain in cultured muscle. Retroviral expression vectors were constructed which encoded the exon-1 sequence of the myosin light chain-1 gene in antisense orientation. These vectors were applied to myogenic cells from embryonic chick and quail pectoralis muscle. Colonies expressing antisense RNA were selected in growth medium containing the neomycin analogue G-418, plus 5-bromo-2′-deoxyuridine (BrdU) and triggered to differentiate by removal of the latter. Expression of antisense myosin light chain-1 mRNA impaired muscle development. In the antisense cultures there were more mononucleated cells, fewer and smaller myotubes which had poorly developed myofibrils and high levels of diffusely staining myosin heavy chain, not apparent in control myotubes. Protein synthesis in the myotube cultures was analyzed by35S-methionine labelling and two-dimensional gel electrophoresis. Except for a suppression of ∼80% of myosin light chain-1fsynthesis, the overall pattern of protein synthesis was not altered significantly. These studies suggest that retardation of myosin light chain-1faccumulation inhibits or delays myofibrillogenesis.
