Mehnert Martin, Sommermeyer Franziska, Berger Maren, Kumar Lakshmipathy Sathish, Gauss Robert, Aebi Markus, Jarosch Ernst, Sommer Thomas
Max-Delbrück-Center for Molecular Medicine, 13125 Berlin, Germany.
Institute of Microbiology, Department of Biology, Swiss Federal Institute of Technology Zurich, 8093 Zurich, Switzerland.
Mol Biol Cell. 2015 Jan 15;26(2):185-94. doi: 10.1091/mbc.E14-07-1202. Epub 2014 Nov 26.
Misfolded proteins of the secretory pathway are extracted from the endoplasmic reticulum (ER), polyubiquitylated by a protein complex termed the Hmg-CoA reductase degradation ligase (HRD-ligase), and degraded by cytosolic 26S proteasomes. This process is termed ER-associated protein degradation (ERAD). We previously showed that the membrane protein Der1, which is a subunit of the HRD-ligase, is involved in the export of aberrant polypeptides from the ER. Unexpectedly, we also uncovered a close spatial proximity of Der1 and the substrate receptor Hrd3 in the ER lumen. We report here on a mutant Hrd3KR that is selectively defective for ERAD of soluble proteins. Hrd3KR displays subtle structural changes that affect its positioning toward Der1. Furthermore, increased quantities of the ER-resident Hsp70-type chaperone Kar2 and the Hsp40-type cochaperone Scj1 bind to Hrd3KR. Of note, deletion of SCJ1 impairs ERAD of model substrates and causes the accumulation of client proteins at Hrd3. Our data imply a function of Scj1 in the removal of malfolded proteins from the receptor Hrd3, which facilitates their delivery to downstream-acting components like Der1.
分泌途径中错误折叠的蛋白质从内质网(ER)中提取出来,被一种称为Hmg-CoA还原酶降解连接酶(HRD连接酶)的蛋白质复合物多聚泛素化,然后由胞质26S蛋白酶体降解。这个过程称为内质网相关蛋白降解(ERAD)。我们之前表明,作为HRD连接酶亚基的膜蛋白Der1参与异常多肽从内质网的输出。出乎意料的是,我们还发现Der1与内质网腔中的底物受体Hrd3在空间上紧密相邻。我们在此报告一种突变体Hrd3KR,它对可溶性蛋白的ERAD有选择性缺陷。Hrd3KR表现出细微的结构变化,影响其与Der1的定位。此外,内质网驻留的Hsp70型伴侣蛋白Kar2和Hsp40型共伴侣蛋白Scj1的数量增加与Hrd3KR结合。值得注意的是,Scj1的缺失会损害模型底物的ERAD,并导致客户蛋白在Hrd3处积累。我们的数据表明Scj1在从受体Hrd3去除错误折叠的蛋白质中发挥作用,这有助于将它们递送至下游作用成分,如Der1。