Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125.
Howard Hughes Medical Institute, California Institute of Technology, Pasadena, CA 91125.
Proc Natl Acad Sci U S A. 2018 Feb 13;115(7):E1455-E1464. doi: 10.1073/pnas.1711218115. Epub 2018 Jan 18.
Cell division in many eukaryotes is driven by a ring containing actin and myosin. While much is known about the main proteins involved, the precise arrangement of actin filaments within the contractile machinery, and how force is transmitted to the membrane, remains unclear. Here we use cryosectioning and cryofocused ion beam milling to gain access to cryopreserved actomyosin rings in for direct 3D imaging by electron cryotomography. Our results show that straight, overlapping actin filaments, running nearly parallel to each other and to the membrane, form a loose bundle of ∼150 nm in diameter that "saddles" the inward-bending membrane at the leading edge of the division septum. The filaments do not make direct contact with the membrane. Our analysis of the actin filaments reveals the variability in filament number, nearest-neighbor distances between filaments within the bundle, their distance from the membrane, and angular distribution with respect to the membrane.
在许多真核生物中,细胞分裂是由含有肌动蛋白和肌球蛋白的环驱动的。虽然已经了解了主要涉及的蛋白质,但对于收缩机制中肌动蛋白丝的精确排列以及力如何传递到膜上,仍然不清楚。在这里,我们使用冷冻切片和冷冻聚焦离子束铣削来获得冷冻保存的肌动球蛋白环,以便通过电子 cryotomography 进行直接的 3D 成像。我们的结果表明,几乎相互平行且与膜平行的直、重叠的肌动蛋白丝形成一个直径约 150nm 的松散束,在分裂隔膜的前缘将向内弯曲的膜“鞍形”。这些细丝与膜没有直接接触。我们对肌动蛋白丝的分析揭示了丝数、束内丝之间的最近邻距离、它们与膜的距离以及相对于膜的角分布的可变性。