Basu Roshni, Munteanu Emilia Laura, Chang Fred
Department of Microbiology and Immunology, Columbia University College of Physicians and Surgeons, New York, NY 10032.
Mol Biol Cell. 2014 Mar;25(5):679-87. doi: 10.1091/mbc.E13-10-0618. Epub 2014 Jan 8.
Yeast and other walled cells possess high internal turgor pressure that allows them to grow and survive in the environment. This turgor pressure, however, may oppose the invagination of the plasma membrane needed for endocytosis. Here we study the effects of turgor pressure on endocytosis in the fission yeast Schizosaccharomyces pombe by time-lapse imaging of individual endocytic sites. Decreasing effective turgor pressure by addition of sorbitol to the media significantly accelerates early steps in the endocytic process before actin assembly and membrane ingression but does not affect the velocity or depth of ingression of the endocytic pit in wild-type cells. Sorbitol also rescues endocytic ingression defects of certain endocytic mutants and of cells treated with a low dose of the actin inhibitor latrunculin A. Endocytosis proceeds after removal of the cell wall, suggesting that the cell wall does not contribute mechanically to this process. These studies suggest that endocytosis is governed by a mechanical balance between local actin-dependent inward forces and opposing forces from high internal turgor pressure on the plasma membrane.
酵母和其他有细胞壁的细胞具有较高的内部膨压,这使它们能够在环境中生长和存活。然而,这种膨压可能会阻碍内吞作用所需的质膜内陷。在这里,我们通过对单个内吞位点进行延时成像,研究了膨压对裂殖酵母粟酒裂殖酵母内吞作用的影响。通过向培养基中添加山梨醇来降低有效膨压,可显著加速肌动蛋白组装和膜内陷之前的内吞过程早期步骤,但不影响野生型细胞内吞小窝的内陷速度或深度。山梨醇还能挽救某些内吞突变体以及用低剂量肌动蛋白抑制剂Latrunculin A处理的细胞的内吞内陷缺陷。去除细胞壁后内吞作用仍可进行,这表明细胞壁在这一过程中没有机械作用。这些研究表明,内吞作用受局部肌动蛋白依赖性内向力与质膜上高内部膨压产生的相反力之间的机械平衡控制。