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On the Role of Myosin-II in Cytokinesis: Division of Dictyostelium Cells under Adhesive and Nonadhesive Conditions

机译:关于肌球蛋白-II在胞质分裂中的作用:在粘附和非粘附条件下盘基网状细胞的分裂

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摘要

We have investigated the role of myosin in cytokinesis in Dictyostelium cells by examining cells under both adhesive and nonadhesive conditions. On an adhesive surface, both wild-type and myosin-null cells undergo the normal processes of mitotic rounding, cell elongation, polar ruffling, furrow ingression, and separation of daughter cells. When cells are denied adhesion through culturing in suspension or on a hydrophobic surface, wild-type cells undergo these same processes. However, cells lacking myosin round up and polar ruffle, but fail to elongate, furrow, or divide. These differences show that cell division can be driven by two mechanisms that we term Cytokinesis A, which requires myosin, and Cytokinesis B, which is cell adhesion dependent. We have used these approaches to examine cells expressing a myosin whose two light chain-binding sites were deleted (ΔBLCBS-myosin). Although this myosin is a slower motor than wild-type myosin and has constitutively high activity due to the abolition of regulation by light-chain phosphorylation, cells expressing ΔBLCBS-myosin were previously shown to divide in suspension ( ). However, we suspected their behavior during cytokinesis to be different from wild-type cells given the large alteration in their myosin. Surprisingly, ΔBLCBS-myosin undergoes relatively normal spatial and temporal changes in localization during mitosis. Furthermore, the rate of furrow progression in cells expressing a ΔBLCBS-myosin is similar to that in wild-type cells.
机译:我们已经通过检查在粘附和非粘附条件下的细胞,研究了肌球蛋白在梭菌细胞中胞质分裂中的作用。在粘附表面上,野生型和肌球蛋白无效的细胞均经历正常的有丝分裂取整,细胞伸长,极谱,皱纹进入和子细胞分离的过程。当通过悬浮培养或在疏水表面上培养而使细胞失去粘附力时,野生型细胞会经历这些相同的过程。然而,缺乏肌球蛋白的细胞会向上聚集和极起皱纹,但不能伸长,犁沟或分裂。这些差异表明,细胞分裂可以由两种机制驱动,我们将这种机制称为胞质分裂A(需要肌球蛋白)和胞质分裂B(依赖细胞粘附)。我们已经使用这些方法来检查表达缺失两个轻链结合位点的肌球蛋白的细胞(ΔBLCBS-肌球蛋白)。尽管此肌球蛋白比野生型肌球蛋白慢,并且由于通过轻链磷酸化取消调节而具有组成性高活性,但先前已显示出表达ΔBLCBS-肌球蛋白的细胞在悬浮液中分裂()。但是,由于肌球蛋白的巨大变化,我们怀疑它们在胞质分裂过程中的行为与野生型细胞不同。令人惊讶的是,ΔBLCBS-肌球蛋白在有丝分裂期间经历相对正常的时空定位变化。此外,在表达ΔBLCBS-肌球蛋白的细胞中犁沟进展的速率与野生型细胞中的相似。

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