Harper J D, McCurdy D W, Sanders M A, Salisbury J L, John P C
Plant Cell Biology Group, Research School of Biological Sciences, Australian National University, Canberra.
Cell Motil Cytoskeleton. 1992;22(2):117-26. doi: 10.1002/cm.970220205.
We have used two monoclonal antibodies to demonstrate the presence and localization of actin in interphase and mitotic vegetative cells of the green alga Chlamydomonas reinhardtii. Commercially available monoclonal antibodies raised against smooth muscle actin (Lessard: Cell Motil. Cytoskeleton 10:349-362, 1988; Lin: Proc. Natl. Acad. Sci. USA 78:2335-2339, 1981) identify Chlamydomonas actin as a approximately 43,000-M(r) protein by Western immunoblot procedures. In an earlier study, Detmers and coworkers (Cell Motil. 5:415-430, 1985) first identified Chlamydomonas actin using NBD-phallacidin and an antibody raised against Dictyostelium actin; they demonstrated that F-actin is localized in the fertilization tubule of mating gametes. Here, we show by immunofluorescence that vegetative Chlamydomonas cells have an array of actin that surrounds the nucleus in interphase cells and undergoes dramatic reorganization during mitosis and cytokinesis. This includes the following: reorganization of actin to the anterior of the cell during preprophase; the formation of a cruciate actin band in prophase; reorganization to a single anterior actin band in metaphase; rearrangement forming a focus of actin anterior to the metaphase plate; reextension of the actin band in anaphase; presence of actin in the forming cleavage furrow during telophase and cytokinesis; and finally reestablishment of the interphase actin array. The studies presented here do not allow us to discriminate between G and F-actin. None the less, our observations, demonstrating dynamic reorganization of actin during the cell cycle, suggest a role for actin that may include the movement of basal bodies toward the spindle poles in mitosis and the formation of the cleavage furrow during cytokinesis.
我们使用了两种单克隆抗体来证明肌动蛋白在莱茵衣藻有丝分裂间期和有丝分裂营养细胞中的存在和定位。通过针对平滑肌肌动蛋白产生的市售单克隆抗体(Lessard:《细胞运动与细胞骨架》10:349 - 362,1988;Lin:《美国国家科学院院刊》78:2335 - 2339,1981),通过蛋白质免疫印迹法将衣藻肌动蛋白鉴定为一种分子量约为43,000的蛋白质。在早期研究中,Detmers及其同事(《细胞运动》5:415 - 430,1985)首次使用NBD - 鬼笔环肽和针对盘基网柄菌肌动蛋白产生的抗体鉴定了衣藻肌动蛋白;他们证明F - 肌动蛋白定位于交配配子的受精管中。在这里,我们通过免疫荧光显示,营养型衣藻细胞有一系列肌动蛋白,在间期细胞中围绕细胞核排列,并在有丝分裂和胞质分裂过程中经历剧烈的重组。这包括以下方面:在前期之前肌动蛋白重新组织到细胞前部;在前期形成十字形肌动蛋白带;在中期重新组织成单个前部肌动蛋白带;重新排列形成中期板前部的肌动蛋白焦点;在后期肌动蛋白带重新延伸;在末期和胞质分裂期间形成的分裂沟中有肌动蛋白存在;最后重新建立间期肌动蛋白阵列。这里呈现的研究不允许我们区分G - 肌动蛋白和F - 肌动蛋白。尽管如此,我们的观察结果表明细胞周期中肌动蛋白的动态重组,提示肌动蛋白可能具有包括在有丝分裂期间基体向纺锤极移动以及在胞质分裂期间形成分裂沟等作用。