Analysis of signal transduction pathways in macrophages using expression vectors with CMV promoters: a cautionary tale.

Madhuri Ramanathan, György Haskó, Samuel Joseph Leibovich

Research output: Contribution to journalArticle

Abstract

The cytomegalovirus (CMV) major immediate-early promoter is a strong promoter used for both in vitro and in vivo expression of proteins in signal transduction and gene therapy studies. CMV activity is induced by external stimuli such as endotoxin from Gram-negative bacteria (LPS), TNF-alpha and phorbol esters. This inducibility poses problems when this promoter is used to drive the expression of either wild type or dominant negative mutated proteins as tools in signal transduction studies. This report draws attention to the problem associated with this widely used approach. The role of NF-kappaB and Hypoxia Inducible Factor-1alpha (HIF-1alpha) in the transcriptional regulation of Vascular Endothelial Growth Factor (VEGF) in macrophages was investigated using CMV-promoter-driven expression of either wild type or dominant negative proteins involved in these pathways. Difficulties encountered while interpreting the data due to the inducibility of the CMV promoter by LPS are highlighted in this report and provide a cautionary note for the evaluation of data acquired using this approach.

Original languageEnglish
Pages (from-to)94-102
Number of pages9
JournalInflammation
Volume29
Issue number2-3
Publication statusPublished - Apr 2005

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Cytomegalovirus
Signal Transduction
Macrophages
Proteins
NF-kappa B
Phorbol Esters
Gram-Negative Bacteria
Endotoxins
Genetic Therapy
Vascular Endothelial Growth Factor A
Tumor Necrosis Factor-alpha

ASJC Scopus subject areas

  • Medicine(all)
  • Immunology
  • Cell Biology

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Analysis of signal transduction pathways in macrophages using expression vectors with CMV promoters : a cautionary tale. / Ramanathan, Madhuri; Haskó, György; Leibovich, Samuel Joseph.

In: Inflammation, Vol. 29, No. 2-3, 04.2005, p. 94-102.

Research output: Contribution to journalArticle

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