出版社：American Society for Biochemistry and Molecular Biology
摘要：Transcriptional profiles of Caenorhabditis elegans grown onunmethylated sterols (desMSs) or on 4-methylsterols (4MSs) werecompared using microarrays. Thirty-four genes were upregulatedand 2 were downregulated >2-fold by growth on 4MSs, including13 cuticle collagen (col) genes, 1 cuticulin gene (cut-1), 2groundhog-like (grl) genes, and 1 groundhog gene (grd-4); col-36and grl-20 were increased 12- and 19-fold, respectively. Fifteenof these 17 genes have been assigned to metabolic mountain 17,suggesting coordinate 4MS-mediated regulation of expression.Quantitative RT-PCR was performed on 27–51 h old animalsgrown on cholesterol (a desMS) or lophenol (a 4MS). col-36 andgrl-20 showed similar cyclic peaks of expression in cholesteroland similar alterations in lophenol, suggesting coregulation.Of six additional grl genes, only grl-3 was upregulated on lophenol;the rest were downregulated. Cyclicity of expression was lostor altered in all six. Nuclear receptor genes nhr-23, nhr-25,nhr-41, and daf-12 all showed cyclic expression in cholesteroland significant downregulation in lophenol by RT-PCR. Expressionof the insulin-like receptor daf-2 was lower in lophenol, whereasthat of its major downstream target daf-16 was higher. Thus,major changes in gene expression accompany growth on 4MSs, butwith surprisingly little effect on normal growth and development.Supplementary key words transcription • reverse transcription polymerase chain reaction • microarray • cholesterol • lathosterol • lophenol • 4-methyl-5-cholest-8(14)-en-3ß-ol • larval development
-methylsterols (4MSs) were compared using microarrays. Thirty-four genes were upregulated and 2 were downregulated >2-fold by growth on 4MSs, including 13 cuticle collagen (col) genes, 1 cuticulin gene (cut-1), 2 groundhog-like (grl) genes, and 1 groundhog gene (grd-4); col-36 and grl-20 were increased 12- and 19-fold, respectively. Fifteen of these 17 genes have been assigned to metabolic mountain 17, suggesting coordinate 4MS-mediated regulation of expression. Quantitative RT-PCR was performed on 27–51 h old animals grown on cholesterol (a desMS) or lophenol (a 4MS). col-36 and grl-20 showed similar cyclic peaks of expression in cholesterol and similar alterations in lophenol, suggesting coregulation. Of six additional grl genes, only grl-3 was upregulated on lophenol; the rest were downregulated. Cyclicity of expression was lost or altered in all six. Nuclear receptor genes nhr-23, nhr-25, nhr-41, and daf-12 all showed cyclic expression in cholesterol and significant downregulation in lophenol by RT-PCR. Expression of the insulin-like receptor daf-2 was lower in lophenol, whereas that of its major downstream target daf-16 was higher. Thus, major changes in gene expression accompany growth on 4MSs, but with surprisingly little effect on normal growth and development.
Supplementary key words transcription • reverse transcription polymerase chain reaction • microarray • cholesterol • lathosterol • lophenol • 4
