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Published online 23 August 2004. doi:10.1083/jcb.200402136
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 166, Number 5, 661-672
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Article

 Calcium-dependent regulation of the cell cycle via a novel MAPK–NF-{kappa}B pathway in Swiss 3T3 cells

Violaine Sée, Nina K.M. Rajala, David G. Spiller, and Michael R.H. White

Centre for Cell Imaging, School of Biological Sciences, University of Liverpool, Liverpool L69 7ZB, England, UK

Address correspondence to M.R.H. White, Centre for Cell Imaging, School of Biological Sciences, University of Liverpool, Liverpool L69 7ZB, England, UK. Tel.: (44) 151-795-4424. Fax: (44) 151-795-4404. email: mwhite{at}liv.ac.uk

Nuclear factor kappa B (NF-{kappa}B) has been implicated in the regulation of cell proliferation and transformation. We investigated the role of the serum-induced intracellular calcium increase in the NF-{kappa}B–dependent cell cycle progression in Swiss 3T3 fibroblasts. Noninvasive photoactivation of a calcium chelator (Diazo-2) was used to specifically disrupt the transient rise in calcium induced by serum stimulation of starved Swiss 3T3 cells. The serum-induced intracellular calcium peak was essential for subsequent NF-{kappa}B activation (measured by real-time imaging of the dynamic p65 and I{kappa}B{alpha} fluorescent fusion proteins), cyclin D1 (CD1) promoter-directed transcription (measured by real-time luminescence imaging of CD1 promoter-directed firefly luciferase activity), and progression to cell division. We further showed that the serum-induced mitogen-activated protein kinase (MAPK) phosphorylation is calcium dependent. Inhibition of the MAPK- but not the PtdIns3K-dependent pathway inhibited NF-{kappa}B signaling, and further, CD1 transcription and cell cycle progression. These data suggest that a serum-dependent calcium signal regulates the cell cycle via a MAPK–NF-{kappa}B pathway in Swiss 3T3 cells.

Key Words: calcium; NF-{kappa}B; MAPK; cyclin D1; flash photolysis

Abbreviations used in this paper: [Ca2+]i, intracellular free calcium concentration; CaMK, Ca-calmodulin kinase; CD1, cyclin D1; NF-{kappa}B, nuclear factor kappa B.


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