Translocation-arrested apolipoprotein B evades proteasome degradation via a sterol-sensitive block in ubiquitin conjugation.
In this study, we explored how sterol metabolism altered by the expression of cholesterol-7alpha-hydroxylase NADPH:oxygen oxidoreductase (7alpha-hydroxylase) affects the ubiquitin-dependent proteasome degradation of translocation-arrested apoB53 in Chinese hamster ovary cells. Stable expression of two different plasmids that encode either rat or human 7alpha-hydroxylase inhibited the ubiquitin conjugation of apoB and its subsequent degradation by the proteasome. Oxysterols (25-hydroxycholesterol and 7-ketocholesterol) reversed the inhibition of apoB degradation caused by 7alpha-hydroxylase. The combined results suggest that the normally rapid proteasome degradation of translocation-arrested apoB can be regulated by a sterol-sensitive polyubiquitin conjugation step in the endoplasmic reticulum. Blocked ubiquitin-dependent proteasome degradation caused translocation-arrested apoB to become sequestered in segregated membrane domains. Our results described for the first time a novel mechanism through which the "quality control" proteasome endoplasmic reticulum degradative pathway of translocation-arrested apoB is linked to sterol metabolism. Sterol-sensitive blocked ubiquitin conjugation appears to selectively inhibit the proteasome degradation of apoB, but not 7alpha-hydroxylase protein, with no impairment of cell vitality or function. Our findings may help to explain why the hepatic production of lipoproteins is increased when familial hypertriglyceridemic patients are treated with drugs that activate 7alpha-hydroxylase (e.g. bile acid-binding resins).
Du
E Z
EZ
Mammalian Cell and Molecular Biology Laboratory, Department of Biology and Molecular Biology Institute, San Diego State University, San Diego, California 92182-0057, USA.
Fleming
J F
JF
Wang
S L
SL
Spitsen
G M
GM
Davis
R A
RA
D000818:Animals; D001055:Apolipoproteins B / Q000378:metabolism*; D001692:Biological Transport; D016466:CHO Cells; D002352:Carrier Proteins / Q000378:metabolism; D002478:Cells, Cultured; D002790:Cholesterol 7-alpha-Hydroxylase / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D006888:Hydroxycholesterols / Q000494:pharmacology; D007976:Leupeptins / Q000378:metabolism; D008861:Microsomes / Q000378:metabolism; D009097:Multienzyme Complexes / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D014162:Transfection; D014452:Ubiquitins / Q000378:metabolism*
D001055:Apolipoproteins B; D002352:Carrier Proteins; D015853:Cysteine Proteinase Inhibitors; D006888:Hydroxycholesterols; D007976:Leupeptins; D009097:Multienzyme Complexes; D012333:RNA, Messenger; D014452:Ubiquitins; C088653:apolipoprotein B-53; C106092:microsomal triglyceride transfer protein; C063261:acetylleucyl-leucyl-norleucinal; C007997:25-hydroxycholesterol; D002790:Cholesterol 7-alpha-Hydroxylase; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex