TY - JOUR
T1 - Molecular identification of the sterol-regulated luminal protease that cleaves SREBPs and controls lipid composition of animal cells
AU - Sakai, Juro
AU - Rawson, Robert B.
AU - Espenshade, Peter J.
AU - Cheng, Dong
AU - Seegmiller, Adam C.
AU - Goldstein, Joseph L.
AU - Brown, Michael S.
N1 - Funding Information:
We thank our colleague Kip Guy for helpful discussions, Lisa Beatty and Vinnie Choudhry for invaluable help with tissue culture, Tammy Dinh and Susan Burke for excellent technical assistance, and Jeff Cormier and Michelle Laremore for DNA sequencing. This work was supported by grants from the National Institutes of Health (NIH) (HL-20948) and the Perot Family Foundation. P. J. E. is the recipient of a NIH Research Science Fellowship Award (HL09993). A. C. S. is supported by NIH Medical Scientist Training Grant GM08014.
PY - 1998/10
Y1 - 1998/10
N2 - The lipid composition of animal cells is controlled by SREBPs, transcription factors released from membranes by sterol-regulated proteolysis. Release is initiated by Site-1 protease (S1P), which cleaves SREBPs in the ER luminal loop between two membrane-spanning regions. To clone S1P, we prepared pCMV-PLAP-BP2, which encodes a fusion protein that contains placental alkaline phosphatase (PLAP) in the ER lumen flanked by cleavage sites for signal peptidase and S1P. In sterol-deprived cells, cleavage by both proteases leads to PLAP secretion. PLAP is not secreted by SRD-12B cells, cholesterol auxotrophs that lack S1P. We transfected SRD-12B cells with pCMV-PLAP-BP2 plus pools of CHO cDNAs and identified a cDNA that restores Site-1 cleavage and PLAP secretion. The cDNA encodes S1P, an intraluminal 1052-amino-acid membrane-bound subtilisin-like protease. We propose that S1P is the sterol-regulated protease that controls lipid metabolism in animal cells.
AB - The lipid composition of animal cells is controlled by SREBPs, transcription factors released from membranes by sterol-regulated proteolysis. Release is initiated by Site-1 protease (S1P), which cleaves SREBPs in the ER luminal loop between two membrane-spanning regions. To clone S1P, we prepared pCMV-PLAP-BP2, which encodes a fusion protein that contains placental alkaline phosphatase (PLAP) in the ER lumen flanked by cleavage sites for signal peptidase and S1P. In sterol-deprived cells, cleavage by both proteases leads to PLAP secretion. PLAP is not secreted by SRD-12B cells, cholesterol auxotrophs that lack S1P. We transfected SRD-12B cells with pCMV-PLAP-BP2 plus pools of CHO cDNAs and identified a cDNA that restores Site-1 cleavage and PLAP secretion. The cDNA encodes S1P, an intraluminal 1052-amino-acid membrane-bound subtilisin-like protease. We propose that S1P is the sterol-regulated protease that controls lipid metabolism in animal cells.
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U2 - 10.1016/S1097-2765(00)80150-1
DO - 10.1016/S1097-2765(00)80150-1
M3 - Article
C2 - 9809072
AN - SCOPUS:0032185770
SN - 1097-2765
VL - 2
SP - 505
EP - 514
JO - Molecular Cell
JF - Molecular Cell
IS - 4
ER -