TY - JOUR
T1 - Critical role of diacylglycerol- and phospholipid-regulated protein kinase Cε in Induction of low-density lipoprotein receptor transcription in response to depletion of cholesterol
AU - Mehta, Kamal D.
AU - Radominska-Pandya, Anna
AU - Kapoor, Gurpreet S.
AU - Dave, Bhuvanesh
AU - Atkins, Brett A.
PY - 2002
Y1 - 2002
N2 - Induction of low-density lipoprotein (LDL) receptor transcription in response to depletion of cellular sterols in animal cells is well established. The intracellular signal or signals involved in regulating this process, however, remain unknown. Using a specific inhibitor of protein kinase C (PKC), calphostin C, we show the requirement of this kinase in the induction process in human hepatoma HepG2 cells. Overexpression of PKCε, but not PKCα, -γ, -δ, or ζ was found to dramatically induce (approximately 18-fold) LDL receptor promoter activity. Interestingly, PKCε-mediated induction was found to be sterol resistant. To further establish that PKCε is involved in the sterol regulation of LDL receptor gene transcription, endogenous PKCε was specifically inhibited by transfection with antisense PKCε phosphorothionate oligonucleotides. Antisense treatment decreased endogenous PKCε protein levels and completely blocked induction of LDL receptor transcription following sterol depletion. PKCε-induced LDL receptor transcription is independent of the extracellular signal-regulated kinase 1 and 2 (p42/44MAPK) cascade, because the MEK-1/2 inhibitor, PD98059 did not inhibit, even though it blocked p42/44MAPK activation. Finally, photoaffinity labeling studies showed an isoform-specific interaction between PKCε and sterols, suggesting that sterols may directly modulate its function by hampering binding of activators. This was confirmed by PKC activity assays. Altogether, these results define a novel signaling pathway leading to induction of LDL receptor transcription following sterol depletion, and a model is proposed to account for a new function for PKCε as part of a sterol-sensitive signal transduction pathway in hepatic cells.
AB - Induction of low-density lipoprotein (LDL) receptor transcription in response to depletion of cellular sterols in animal cells is well established. The intracellular signal or signals involved in regulating this process, however, remain unknown. Using a specific inhibitor of protein kinase C (PKC), calphostin C, we show the requirement of this kinase in the induction process in human hepatoma HepG2 cells. Overexpression of PKCε, but not PKCα, -γ, -δ, or ζ was found to dramatically induce (approximately 18-fold) LDL receptor promoter activity. Interestingly, PKCε-mediated induction was found to be sterol resistant. To further establish that PKCε is involved in the sterol regulation of LDL receptor gene transcription, endogenous PKCε was specifically inhibited by transfection with antisense PKCε phosphorothionate oligonucleotides. Antisense treatment decreased endogenous PKCε protein levels and completely blocked induction of LDL receptor transcription following sterol depletion. PKCε-induced LDL receptor transcription is independent of the extracellular signal-regulated kinase 1 and 2 (p42/44MAPK) cascade, because the MEK-1/2 inhibitor, PD98059 did not inhibit, even though it blocked p42/44MAPK activation. Finally, photoaffinity labeling studies showed an isoform-specific interaction between PKCε and sterols, suggesting that sterols may directly modulate its function by hampering binding of activators. This was confirmed by PKC activity assays. Altogether, these results define a novel signaling pathway leading to induction of LDL receptor transcription following sterol depletion, and a model is proposed to account for a new function for PKCε as part of a sterol-sensitive signal transduction pathway in hepatic cells.
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U2 - 10.1128/MCB.22.11.3783-3793.2002
DO - 10.1128/MCB.22.11.3783-3793.2002
M3 - Article
C2 - 11997513
AN - SCOPUS:0036094341
VL - 22
SP - 3783
EP - 3793
JO - Molecular and Cellular Biology
JF - Molecular and Cellular Biology
SN - 0270-7306
IS - 11
ER -