Kiel Jan A K W, Emmrich Kerstin, Meyer Helmut E, Kunau Wolf-H
Abteilung für Zellbiochemie, Medizinische Fakultät der Ruhr-Universität Bochum, Germany.
J Biol Chem. 2005 Jan 21;280(3):1921-30. doi: 10.1074/jbc.M403632200. Epub 2004 Nov 9.
PEX genes encode proteins (peroxins) that are required for the biogenesis of peroxisomes. One of these peroxins, Pex5p, is the receptor for matrix proteins with a type 1 peroxisomal targeting signal (PTS1), which shuttles newly synthesized proteins from the cytosol into the peroxisome matrix. We observed that in various Saccharomyces cerevisiae pex mutants disturbed in the early stages of PTS1 import, the steady-state levels of Pex5p are enhanced relative to wild type controls. Furthermore, we identified ubiquitinated forms of Pex5p in deletion mutants of those PEX genes that have been implicated in recycling of Pex5p from the peroxisomal membrane into the cytosol. Pex5p ubiquitination required the presence of the ubiquitin-conjugating enzyme Ubc4p and the peroxins that are required during early stages of PTS1 protein import. Finally, we provide evidence that the proteasome is involved in the turnover of Pex5p in wild type yeast cells, a process that requires Ubc4p and occurs at the peroxisomal membrane. Our data suggest that during receptor recycling a portion of Pex5p becomes ubiquitinated and degraded by the proteasome. We propose that this process represents a conserved quality control mechanism in peroxisome biogenesis.
PEX基因编码过氧化物酶体生物发生所需的蛋白质(过氧化物酶)。其中一种过氧化物酶Pex5p是具有1型过氧化物酶体靶向信号(PTS1)的基质蛋白的受体,它将新合成的蛋白质从细胞质转运到过氧化物酶体基质中。我们观察到,在PTS1导入早期受到干扰的各种酿酒酵母pex突变体中,Pex5p的稳态水平相对于野生型对照有所提高。此外,我们在那些与Pex5p从过氧化物酶体膜循环到细胞质中有关的PEX基因的缺失突变体中鉴定出了泛素化形式的Pex5p。Pex5p泛素化需要泛素结合酶Ubc4p和PTS1蛋白导入早期所需的过氧化物酶。最后,我们提供证据表明蛋白酶体参与野生型酵母细胞中Pex5p的周转,这一过程需要Ubc4p且发生在过氧化物酶体膜上。我们的数据表明,在受体循环过程中,一部分Pex5p会被泛素化并被蛋白酶体降解。我们提出,这一过程代表了过氧化物酶体生物发生中一种保守的质量控制机制。