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Papers In Press, published online ahead of print November 1, 2003
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Journal of Lipid Research, Vol. 44, 2109-2119, November 2003
Copyright © 2003 by American Society for Biochemistry and Molecular Biology
Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University, 1230 York Avenue, Box 179, New York, NY 10021
1 To whom correspondence should be addressed. e-mail: breslow{at}rockefeller.edu
High-cholesterol diets elicit changes in gene expression via such transcription factors as sterol-regulatory element binding proteins (SREBPs) and liver X receptors (LXRs). We used Affymetrix microarrays to identify genes in mouse liver regulated by dietary cholesterol (0.0% vs. 0.5% cholesterol wt/wt). Three independent experiments were performed, and data were analyzed with Affymetrix Microarray Suite and ANOVA statistical software. There were 69 unique Unigene clusters consistently regulated by dietary cholesterol (37 downregulated and 32 upregulated). The array results were confirmed by quantitative RT-PCR (Q-PCR) for seven of nine downregulated genes and five of six upregulated genes. A time course of dietary cholesterol feeding over 1 week revealed different temporal patterns of gene regulation for these confirmed genes. Six downregulated genes were examined in transgenic mice overexpressing truncated nuclear forms of SREBP-1a and SREBP-2, and all were induced in these mice. A second microarray analysis of mice treated with the LXR agonist TO901317 confirmed that 13 of the 32 cholesterol upregulated genes were also LXR-activated. This array result was confirmed by Q-PCR for three of three genes.
In summary, these studies identified and confirmed six novel dietary cholesterol-regulated genes, three putative SREBP target genes (calcium/calmodulin-dependent protein kinase 1D, fatty acid binding protein 5, and proprotein convertase subtilisin/kexin 9), and three putative LXR target genes (a disintegrin and metalloprotease domain 11, apoptosis-inhibitory 6, and F-box-only protein 3).
Abbreviations: Abcg5, ATP binding cassette, subfamily G, member 5; Acac, acetyl-CoA carboxylase; Acly, ATP citrate lyase; Adam11, a disintegrin and metalloprotease domain 11; Api6, apoptosis-inhibitory 6; Camk1D, calcium/calmodulin-dependent protein kinase 1D; CYP7A1, cholesterol 7-
-hydroxylase; FABP5, fatty acid binding protein 5; Fbxo3, F-box-only protein 3; HMGCR, HMG-CoA reductase; HMGCS, HMG-CoA synthase; Laptm5, lysosomal-associated protein transmembrane 5; LXR, liver X receptor; Pcsk9, proprotein convertase subtilisin/kexin 9; Rgs16, regulator of G-protein signaling 16; SAA3, serum amyloid A 3; Sqle, squalene epoxidase; SREBP, sterol-regulatory element binding protein; StarD4, StAR-related lipid transfer domain containing 4
Supplementary key words dietary cholesterol • oligonucleotide arrays • gene expression profiling • sterol-regulatory element binding proteins • liver X receptor
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