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J. Lipid Res.
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Originally published In Press as doi:10.1194/jlr.M800259-JLR200 on June 26, 2008

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Journal of Lipid Research, Vol. 49, 2197-2211, October 2008
Copyright © 2008 by American Society for Biochemistry and Molecular Biology

Stimulation of the human CTP:phosphoethanolamine cytidylyltransferase gene by early growth response protein 1

Lin Zhu, Christa Johnson and Marica Bakovic1

Department of Human Health and Nutritional Sciences, University of Guelph, Guelph, Ontario, Canada N1G 2W1

This work was supported by a Canadian Institute of Health Research Scholarship (C.J.) and operating Grants MOP-68962 and 410-2007-86448 from the Canadian Institute of Health Research (M.B.).

Published, JLR Papers in Press, June 26, 2008.

1 To whom correspondence should be addressed. e-mail: mbakovic{at}uoguelph.ca

Change in phosphoethanolamine pool size in tumor tissues is an important indicator of tumor prognosis and drug therapy efficacy. Phosphoethanolamine is the substrate of the regulatory enzyme CTP:phosphoethanolamine cytidylyltransferase (ECT) in the de novo biosynthesis of phosphatidylethanolamine (PE). Metabolic labeling with [14C]ethanolamine revealed a reduced ECT activity in MCF-7 breast cancer cells, which led to an accumulation of phosphoethanolamine and a decrease in PE synthesis in comparison with MCF-10A mammary epithelial cells. The enhanced ECT activity in MCF-10A cells was due to significantly elevated CTP:phosphoethanolamine cytidylyltransferase gene (PCYT2) expression, at the level of promoter activity, mRNA, and protein content. The early growth response protein 1 (EGR1) could account for most of the elevated ECT activity in MCF-10A cells relative to MCF-7 cells, as evidenced by promoter-luciferase reporter assays, gel-shift analyses, and by alterations in the EGR1 gene expression. In MCF-7 cells, EGR1 is present at lower levels and the basal PCYT2 promoter activity is maintained by proximal CAAT and GC regions and by elevated nuclear NF{kappa}B activity. Together, these data demonstrate that EGR1 is an important transcriptional stimulator of the human PCYT2 and that conditions that modify EGR1 also affect the function of ECT and consequently PE synthesis.

Supplementary key words PCYT2 • EGR1 • NF{kappa}B • phospholipids • phosphatidylethanolamine


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