Derlin-1 and UBXD8 are engaged in dislocation and degradation of lipidated ApoB-100 at lipid droplets

• Published in: Molecular biology of the cell Vol. 23; no. 5; pp. 800 - 810
• Main Authors: Suzuki, Michitaka, Otsuka, Toshihiko, Ohsaki, Yuki, Cheng, Jinglei, Taniguchi, Takako, Hashimoto, Hisashi, Taniguchi, Hisaaki, Fujimoto, Toyoshi
• Format: Journal Article
• Language: English
• Published: United States The American Society for Cell Biology 01.03.2012
• Subjects: Animals; Apolipoprotein B-100 - metabolism; Blood Proteins - genetics; Blood Proteins - metabolism; Cell Line, Tumor; Chlorocebus aethiops; COS Cells; Cytoplasm - metabolism; Endoplasmic Reticulum-Associated Degradation; HeLa Cells; Humans; Membrane Proteins - genetics; Membrane Proteins - metabolism; Proteolysis; RNA, Small Interfering - genetics
• ISSN: 1059-1524; 1939-4586
• DOI: 10.1091/mbc.e11-11-0950

Apolipoprotein B-100 (ApoB) is the principal component of very low density lipoprotein. Poorly lipidated nascent ApoB is extracted from the Sec61 translocon and degraded by proteasomes. ApoB lipidated in the endoplasmic reticulum (ER) lumen is also subjected to proteasomal degradation, but where and how it dislocates to the cytoplasm remain unknown. In the present study, we demonstrate that ApoB after lipidation is dislocated to the cytoplasmic surface of lipid droplets (LDs) and accumulates as ubiquitinated ApoB in Huh7 cells. Depletion of UBXD8, which is almost confined to LDs in this cell type, decreases recruitment of p97 to LDs and causes an increase of both ubiquitinated ApoB on the LD surface and lipidated ApoB in the ER lumen. In contrast, abrogation of Derlin-1 function induces an accumulation of lipidated ApoB in the ER lumen but does not increase ubiquitinated ApoB on the LD surface. UBXD8 and Derlin-1 bind with each other and with lipidated ApoB and show colocalization around LDs. These results indicate that ApoB after lipidation is dislocated from the ER lumen to the LD surface for proteasomal degradation and that Derlin-1 and UBXD8 are engaged in the predislocation and postdislocation steps, respectively.