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Journal of Lipid Research, Vol. 45, 1943-1951, October 2004
Copyright © 2004 by American Society for Biochemistry and Molecular Biology
* Biochemistry, Cell Biology, and Metabolism, Nagoya City University Graduate School of Medical Sciences, Kawasumi 1, Mizuho-cho, Mizuho-ku, Nagoya 467-8601, Japan
Department of Biochemistry, Dartmouth Medical School, Hanover, NH 03755
2 To whom correspondence should be addressed. e-mail: syokoyam{at}med.nagoya-cu.ac.jp
Differential regulation has been suggested for cellular cholesterol and phospholipid release mediated by apolipoprotein A-I (apoA-I)/ABCA1. We investigated various factors involved in cholesterol mobilization related to this pathway. ApoA-I induced a rapid decrease of the cellular cholesterol compartment that is in equilibrium with the ACAT-accessible pool in cells that generate cholesterol-rich HDL. Pharmacological and genetic inactivation of ACAT enhanced the apoA-I-mediated cholesterol release through upregulation of ABCA1 and through cholesterol enrichment in the HDL generated. Pharmacological activation of protein kinase C (PKC) also decreased the ACAT-accessible cholesterol pool, not only in the cells that produce cholesterol-rich HDL by apoA-I (i.e., human fibroblast WI-38 cells) but also in the cells that generate cholesterol-poor HDL (mouse fibroblast L929 cells). In L929 cells, the PKC activation caused an increase in apoA-I-mediated cholesterol release without detectable change in phospholipid release and in ABCA1 expression.
These results indicate that apoA-I mobilizes intracellular cholesterol for the ABCA1-mediated release from the compartment that is under the control of ACAT. The cholesterol mobilization process is presumably related to PKC activation by apoA-I.
Abbreviations: apoA-I, apolipoprotein A-I; DOG, sn-1,2-dioctanoylglycerol; PKC, protein kinase C; PMA, phorbol 12-myristate-13-acetate; SREBP, sterol regulatory element binding protein
Supplementary key words ATP binding cassette transporter A1 • acyl-coenzyme A:cholesterol acyltransferase • apolipoprotein A-I • protein kinase C
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