The Antiapoptotic Gene mcl-1 Is Up-Regulated by the Phosphatidylinositol 3-Kinase/Akt Signaling Pathway through a Transcription Factor Complex Containing CREB

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Molecular and Cellular Biology, September 1999, p. 6195-6206, Vol. 19, No. 9
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The Antiapoptotic Gene mcl-1 Is Up-Regulated by the Phosphatidylinositol 3-Kinase/Akt Signaling Pathway through a Transcription Factor Complex Containing CREB

Ju-Ming Wang,¹^,² Jyh-Rong Chao,²^,³ Wannhsin Chen,¹^,⁴ Min-Liang Kuo,³ Jeffrey J.-Y. Yen,¹^,⁴ and Hsin-Fang Yang-Yen¹^,²^,^*

Graduate Institute of Life Science, National Defense Medical School,¹ Institute of Molecular Biology² and Institute of Biomedical Sciences,⁴ Academia Sinica, and Institute of Toxicology, National Taiwan University Medical School,³ Taipei, Taiwan, Republic of China

Received 16 February 1999/Returned for modification 30 March 1999/Accepted 21 June 1999

mcl-1 is an immediate-early gene activated by the granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin3 (IL-3) signaling pathways and plays an important role in theviability response of these cytokines. In this study, we demonstratedthat cytokine stimulation of mcl-1 mRNA and protein expressionwere attenuated by pretreatment of cells with phosphatidylinositol3-kinase (PI3-K) inhibitors. Reporter gene assays further showedthat the PI3-K/Akt signaling pathway was involved in IL-3 activationof mcl-1 gene transcription. Analysis of the mcl-1 promoter revealedthat both promoter elements, SIE at position $-$ 87 and CRE-2 at $-$ 70, contribute to IL-3 stimulation of mcl-1 gene expression.Although either the SIE site or the CRE-2 site alone was sufficientto confer IL-3 inducibility on a heterologous promoter, only IL-3activation of the CRE-2 reporter was mediated via the PI3-K/Aktpathway. The SIE binding activity was constitutively high in cellsdeprived of or stimulated by IL-3. In contrast, the CRE-2 bindingactivity was low in cytokine-starved cells and was strongly inducedwithin 1 h following cytokine treatment of cells. In addition,cytokine induction of the CRE-2 but not of the SIE binding activitywas dependent on activation of the PI3-K/Akt signaling pathway.Lastly, we showed that CREB was one component of the CRE-2 bindingcomplex and played a role in IL-3 activation of the mcl-1 reportergene. Taken together, our results suggest that both PI3-K/Akt-dependentand -independent pathways contribute to the IL-3 activation ofmcl-1 gene expression. Activation of mcl-1 by the PI3-K/Akt-dependentpathway is through a transcription factor complex containingCREB.

^* Corresponding author. Mailing address: Institute of Molecular Biology, Academia Sinica, 128 Yen-Jiou Yuan Rd. Sec. 2, NangKang,Taipei, 11529 Taiwan, R.O.C. Phone: 886-2-2789-9228. Fax: 886-2-2782-6085.E-mail: IMBYY{at}ccvax.sinica.edu.tw.

Molecular and Cellular Biology, September 1999, p. 6195-6206, Vol. 19, No. 9
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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