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Upregulation of AKT1 protein expression in forskolin-stimulated macrophage: evidence from ChIP analysis that CREB binds to and activates the AKT1 promoter.

Publication ,  Journal Article
Misra, UK; Pizzo, SV
Published in: J Cell Biochem
March 1, 2007

Recently, we reported that silencing CREB gene expression by RNAi significantly attenuates forskolin-induced activation of Akt1. We now provide evidence that forskolin-treatment causes transcriptional and translational upregulation of Akt1 in macrophages. Akt synthesis was demonstrated by [(14)C]leucine or [(35)S] incorporation into newly synthesized Akt1 protein. Akt protein levels increased by approximately 1.5-fold after only a 5 min exposure of macrophages to forskolin. Akt1 levels thereafter rapidly returned to basal values (t(1/2) approximately 15 min). Maximal upregulation of Akt1 occurred in cells treated with 10 microM forskolin. Forskolin-dependent Akt1 synthesis was abolished by pretreating the cells with CREB-directed dsRNA as demonstrated at both the message and protein level, as well as by determining the synthesis of [(35)S]-labeled Akt1 protein. The PKA inhibitor H-89, greatly attenuated forskolin-induced Akt1 synthesis. Transcriptional and translational inhibitors also greatly reduced Akt1 synthesis in forskolin-stimulated [(14)C]leucine-labeled macrophages. Using a chromatin immunoprecipitation assay, we demonstrate that CREB binds to a CRE binding domain of the Akt1 gene promoter. In conclusion, we show here for the first time transcriptional upregulation of Akt1 by CREB, based upon Akt1 protein synthesis and its modulation by transitional and translational inhibitors in forskolin-stimulated cells, Akt1 protein. and mRNA levels upon silencing CREB gene expression, and binding of CREB to the Akt1 gene promoter.

Duke Scholars

Published In

J Cell Biochem

DOI

ISSN

0730-2312

Publication Date

March 1, 2007

Volume

100

Issue

4

Start / End Page

1022 / 1033

Location

United States

Related Subject Headings

  • Up-Regulation
  • Sulfonamides
  • RNA, Messenger
  • RNA Interference
  • Proto-Oncogene Proteins c-akt
  • Protein Binding
  • Promoter Regions, Genetic
  • Models, Biological
  • Mice, Inbred C57BL
  • Mice
 

Citation

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Misra, U. K., & Pizzo, S. V. (2007). Upregulation of AKT1 protein expression in forskolin-stimulated macrophage: evidence from ChIP analysis that CREB binds to and activates the AKT1 promoter. J Cell Biochem, 100(4), 1022–1033. https://doi.org/10.1002/jcb.21086
Misra, Uma Kant, and Salvatore Vincent Pizzo. “Upregulation of AKT1 protein expression in forskolin-stimulated macrophage: evidence from ChIP analysis that CREB binds to and activates the AKT1 promoter.J Cell Biochem 100, no. 4 (March 1, 2007): 1022–33. https://doi.org/10.1002/jcb.21086.
Misra, Uma Kant, and Salvatore Vincent Pizzo. “Upregulation of AKT1 protein expression in forskolin-stimulated macrophage: evidence from ChIP analysis that CREB binds to and activates the AKT1 promoter.J Cell Biochem, vol. 100, no. 4, Mar. 2007, pp. 1022–33. Pubmed, doi:10.1002/jcb.21086.
Journal cover image

Published In

J Cell Biochem

DOI

ISSN

0730-2312

Publication Date

March 1, 2007

Volume

100

Issue

4

Start / End Page

1022 / 1033

Location

United States

Related Subject Headings

  • Up-Regulation
  • Sulfonamides
  • RNA, Messenger
  • RNA Interference
  • Proto-Oncogene Proteins c-akt
  • Protein Binding
  • Promoter Regions, Genetic
  • Models, Biological
  • Mice, Inbred C57BL
  • Mice