ANTISENSE RNA INACTIVATION OF MYOSIN HEAVY-CHAIN GENE-EXPRESSION IN DICTYOSTELIUM-DISCOIDEUM
ANTISENSE RNA INACTIVATION OF MYOSIN HEAVY-CHAIN GENE-EXPRESSION IN DICTYOSTELIUM-DISCOIDEUM
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DOI：
10.1126/science.3576221
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发表时间：
1987-05-29
期刊：
影响因子：
56.9
通讯作者：
LOOMIS, WF
中科院分区：
文献类型：
--
作者：
KNECHT, DA;LOOMIS, WF
肌球蛋白在横纹肌细胞收缩中的作用是众所周知的，但其在非肌肉细胞中的重要性尚不清楚。通过分离特异性缺乏肌球蛋白重链（MHC-A）蛋白的细胞，研究了盘基网柄藻（Dictyosterichdiscoideum）中肌球蛋白的功能。用编码与反义RNA互补的RNA的载体转化细胞。稳定的转化体具有MHC A蛋白量的显著减少，生长缓慢，并产生巨大的多核后代，表明胞质分裂受损。令人惊讶的是，细胞粘附在表面，延伸伪足，并能够变形运动。由饥饿细胞启动的发育序列由于缺乏肌球蛋白而严重受损。这些细胞不能正常形成多细胞聚集体，并且不经历随后的形态发生。通过将食物来源从液体培养基改变为细菌，相对于反义RNA的表达，内源性mRNA的表达可以增加。当以这种方式生长时，转化的细胞积累MHC A蛋白，保持单核，并正常发育。
The role of myosin in the contraction of striated muscle cells is well known, but its importance in nonmuscle cells is not yet clear. The function of myosin inDictyostelium discoideumhas been investigated by isolating cells which specifically lack myosin heavy chain (MHC A) protein. Cells were transformed with a vector encoding RNA complementary tomhcAmessenger RNA (antisense RNA). Stable transformants have a dramatic reduction in the amount of MHC A protein, grow slowly, and generate giant multinucleated progeny, indicating an impairment in cytokinesis. Surprisingly, the cells adhere to surfaces, extend pseudopods and are capable of ameboid locomotion. The developmental sequence that is initiated by starving cells is severely impaired by the lack of myosin. The cells are unable to form multicellular aggregates normally and do not undergo subsequent morphogenesis. By changing the food source from liquid medium to bacteria, expression of the endogenousmhcAmessenger RNA can be increased relative to expression of antisense RNA. When grown in this way, the transformed cells accumulate MHC A protein, remain mononucleate, and proceed through development normally.