Motley Alison M, Ward Gemma P, Hettema Ewald H
Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield, S10 2TN, UK.
J Cell Sci. 2008 May 15;121(Pt 10):1633-40. doi: 10.1242/jcs.026344. Epub 2008 Apr 29.
Yeast peroxisomes multiply by fission. Fission requires two dynamin-related proteins, Dnm1p and Vps1p. Using an in vivo fission assay, we show that Dnm1p-dependent peroxisome fission requires Fis1p, Caf4p and Mdv1p. Fluorescence microscopy of cells expressing GFP-tagged Caf4p and Mdv1p revealed that their association with peroxisomes relies on Fis1p. Vps1p-dependent peroxisome fission occurs independently of these factors. Vps1p contributes most to fission of peroxisomes when cells are grown on glucose. Overexpression of Dnm1p suppresses the fission defect as long as Fis1p and either Mdv1p or Caf4p are present. Conversely, overexpression of Dnm1p does not restore the vacuolar fusion defect of vps1 cells and Vps1p overexpression does not restore the mitochondrial fission defect of dnm1 cells. These data show that Vps1p and Dnm1p are part of independent fission machineries. Because the contribution of Dnm1p to peroxisome fission appears to be more pronounced in cells that proliferate peroxisomes in response to mitochondrial dysfunction, Dnm1p might be part of the mechanism that coordinates mitochondrial and peroxisomal biogenesis.
酵母过氧化物酶体通过分裂增殖。分裂需要两种与发动蛋白相关的蛋白质,即Dnm1p和Vps1p。利用体内分裂检测法,我们发现依赖Dnm1p的过氧化物酶体分裂需要Fis1p、Caf4p和Mdv1p。对表达绿色荧光蛋白标记的Caf4p和Mdv1p的细胞进行荧光显微镜观察发现,它们与过氧化物酶体的结合依赖于Fis1p。依赖Vps1p的过氧化物酶体分裂独立于这些因子发生。当细胞在葡萄糖上生长时,Vps1p对过氧化物酶体分裂的贡献最大。只要存在Fis1p以及Mdv1p或Caf4p其中之一,Dnm1p的过表达就能抑制分裂缺陷。相反,Dnm1p的过表达不能恢复vps1细胞的液泡融合缺陷,Vps1p的过表达也不能恢复dnm1细胞的线粒体分裂缺陷。这些数据表明,Vps1p和Dnm1p是独立分裂机制的组成部分。由于在因线粒体功能障碍而过氧化物酶体增殖的细胞中,Dnm1p对过氧化物酶体分裂的贡献似乎更为显著,所以Dnm1p可能是协调线粒体和过氧化物酶体生物发生机制的一部分。