Effectors of rapid homeostatic responses of endoplasmic reticulum cholesterol and 3-hydroxy-3-methylglutaryl-CoA reductase.
The cholesterol content of the endoplasmic reticulum (ER) and the activity of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) imbedded therein respond homeostatically within minutes to changes in the level of plasma membrane cholesterol. We have now examined the roles of sterol regulatory element-binding protein (SREBP)-dependent gene expression, side chain oxysterol biosynthesis, and cholesterol precursors in the short term regulation of ER cholesterol levels and HMGR activity. We found that SREBP-dependent gene expression is not required for the response to changes in cell cholesterol of either the pool of ER cholesterol or the rate of cholesterol esterification. It was also found that the acute proteolytic inactivation of HMGR triggered by cholesterol loading required the conversion of cholesterol to 27-hydroxycholesterol. High levels of exogenous 24,25-dihydrolanosterol drove the inactivation of HMGR; lanosterol did not. However, purging endogenous 24,25-dihydrolanosterol, lanosterol, and other biosynthetic sterol intermediates by treating cells with NB-598 did not greatly affect either the setting of their ER cholesterol pool or the inactivation of their HMGR. In summary, neither SREBP-regulated genes nor 27-hydroxycholesterol is involved in setting the ER cholesterol pool. On the other hand, 27-hydroxycholesterol, rather than cholesterol itself or biosynthetic precursors of cholesterol, stimulates the rapid inactivation of HMGR in response to high levels of cholesterol.
Lange
Yvonne
Y
Department of Pathology, Rush University Medical Center, Chicago, Illinois 60612. Electronic address: ylange@rush.edu.
Ory
Daniel S
DS
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Ye
Jin
J
Department of Pathology, Rush University Medical Center, Chicago, Illinois 60612.
Lanier
Michael H
MH
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Hsu
Fong-Fu
FF
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Steck
Theodore L
TL
Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, Illinois 60637.
D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D004790:Enzyme Induction / Q000187:drug effects; D004951:Esterification / Q000187:drug effects; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008401:Gas Chromatography-Mass Spectrometry; D006706:Homeostasis / Q000187:drug effects; D006801:Humans; D006903:Hydroxymethylglutaryl CoA Reductases / Q000096:biosynthesis; D007810:Lanosterol / Q000378:metabolism; D007976:Leupeptins / Q000494:pharmacology; D008297:Male; D051779:Sterol Regulatory Element Binding Proteins / Q000378:metabolism; D019294:Xanthomatosis, Cerebrotendinous / Q000201:enzymology / Q000473:pathology
D007976:Leupeptins; D051779:Sterol Regulatory Element Binding Proteins; D007810:Lanosterol; D002784:Cholesterol; D006903:Hydroxymethylglutaryl CoA Reductases; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde