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A more recent version of this article appeared on October 1, 2004

Papers In Press, published online ahead of print August 1, 2004
J. Lipid Res., doi:10.1194/jlr.M400264-JLR200
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Submitted on July 12, 2004
Revised on July 28, 2004
Accepted on August 1, 2004

Intracellular cholesterol mobilization involved in the ABCA1/apolipoprotein-mediated assembly of high density lipoprotein in fibroblasts

Yoshio Yamauchi, Catherine C. Y. Chang, Michi Hayashi, Sumiko Abe-Dohmae, Patrick C. Reid, Ta-Yuan Chang, and Shinji Yokoyama

Biochemistry, Cell Biology and Metabolism, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi 467-8601

Corresponding Author: 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)/ATP-binding cassette transporter A1 (ABCA1). We investigated various factors involved in cholesterol mobilization related to this pathway. ApoA-I induced rapid decrease of the cellular cholesterol compartment that is in equilibrium with the acyl-CoA: cholesterol acyltransferase (ACAT)-accessible pool, in the cells that generate cholesterol-rich HDL. Pharmacological and ge netic 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 cho lesterol pool, not only in the cells that produce cholesterol-rich HDL by apoA-I (i.e. human fibroblast cell WI-38) but also in the cells that generate cholesterol-poor HDL (mouse fibroblast cell L929). In L929 cells, the PKC activation caused increase i n 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 of which availabil ity is under the control of ACAT. The cholesterol mobilization process is presumably related to PKC activation by apoA-I.o


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