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Calyculin A, an enhancer of myosin, speeds up anaphase chromosome movement

机译：Calyculin A，一种肌球蛋白的增强剂，可加速后期染色体的移动

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Actin and myosin inhibitors often blocked anaphase movements in insect spermatocytes in previous experiments. Here we treat cells with an enhancer of myosin, Calyculin A, which inhibits myosin-light-chain phosphatase from dephosphorylating myosin; myosin thus is hyperactivated. Calyculin A causes anaphase crane-fly spermatocyte chromosomes to accelerate poleward; after they reach the poles they often move back toward the equator. When added during metaphase, chromosomes at anaphase move faster than normal. Calyculin A causes prometaphase chromosomes to move rapidly up and back along the spindle axis, and to rotate. Immunofluorescence staining with an antibody against phosphorylated myosin regulatory light chain (p-squash) indicated increased phosphorylation of cleavage furrow myosin compared to control cells, indicating that calyculin A indeed increased myosin phosphorylation. To test whether the Calyculin A effects are due to myosin phosphatase or to type 2 phosphatases, we treated cells with okadaic acid, which inhibits protein phosphatase 2A at concentrations similar to Calyculin A but requires much higher concentrations to inhibit myosin phosphatase. Okadaic acid had no effect on chromosome movement. Backward movements did not require myosin or actin since they were not affected by 2,3-butanedione monoxime or LatruculinB. Calyculin A affects the distribution and organization of spindle microtubules, spindle actin, cortical actin and putative spindle matrix proteins skeletor and titin, as visualized using immunofluorescence. We discuss how accelerated and backwards movements might arise.
机译：在先前的实验中，肌动蛋白和肌球蛋白抑制剂通常会阻止昆虫精母细胞的后期运动。在这里，我们用肌球蛋白增强剂Calyculin A处理细胞，该增强剂抑制肌球蛋白轻链磷酸酶使肌球蛋白去磷酸化。肌球蛋白因此被过度激活。 Calyculin A导致后期白鹤飞行的精母细胞染色体向极加速;到达极点后，他们通常会向赤道移动。当在中期添加时，后期染色体移动得比正常染色体快。 Calyculin A导致前中期染色体沿着纺锤轴快速上下移动并旋转。用抗磷酸化的肌球蛋白调节性轻链（p-squash）抗体进行的免疫荧光染色表明，与对照细胞相比，沟沟肌球蛋白的裂解磷酸化增加，这表明花萼蛋白A确实增加了肌球蛋白的磷酸化。为了测试Calyculin A的作用是由于肌球蛋白磷酸酶还是2型磷酸酶引起的，我们用冈田酸处理了细胞，冈田酸以与Calyculin A相似的浓度抑制蛋白磷酸酶2A，但需要更高的浓度来抑制肌球蛋白磷酸酶。冈田酸对染色体运动没有影响。向后运动不需要肌球蛋白或肌动蛋白，因为它们不受2，3-丁二酮一肟或LatruculinB的影响。 Calyculin A可影响纺锤体微管，纺锤体肌动蛋白，皮层肌动蛋白和假定的纺锤体基质蛋白骨架和纤溶蛋白的分布和组织，如使用免疫荧光法所见。我们讨论了如何加速和向后运动。

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