Abstract
Prolonged and continuous exposure to growth factors is required to commit cells to the cell cycle. Here we show that the prolonged requirement for growth factor can be replaced with two short pulses of mitogen. The first pulse of growth factor moves the cell through the initial segment of the G0 to S interval. This initial pulse also makes cells responsive to a second pulse of growth factor, which engages components of the cell-cycle machinery necessary for progression into S phase. We also show that activation of MAP kinase kinase (MEK) and induction of the transcription factor c-Myc are sufficient to drive the first, but not the second, phase of signalling. Furthermore, synthetic phosphatidylinositol-3-OH kinase (PI(3)K) lipid products are sufficient to drive the second phase of signalling, but not the first. These findings suggest that there is a common signalling cascade by which mitogens drive arrested cells into the cell cycle, and that this cascade involves the temporally coordinated input of MEK, c-Myc and PI(3)K.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Pardee, A. B. A restriction point for control of normal animal cell proliferation. Proc. Natl Acad. Sci. USA 71, 1286– 1290 (1974).
Pardee, A. B. G1 events and regulation of cell proliferation. Science 246, 603–608 (1989).
Planas-Silva, M. D. & Weinberg, R. A. The restriction point and control of cell proliferation. Curr. Opin. Cell Biol. 9, 768–772 ( 1997).
Aktas, H., Cai, H. & Cooper, G. M. Ras links growth factor signaling to the cell cycle machinery via regulation of cyclin D1 and the Cdk inhibitor p27KIP1. Mol. Cell. Biol. 17, 3850–3857 (1997).
Winston, J. T., Coats, S. R., Wang, Y. Z. & Pledger, W. J. Regulation of the cell cycle machinery by oncogenic ras. Oncogene 12, 127–134 ( 1996).
Kerkhoff, E. & Rapp, U. R. Induction of cell proliferation in quiescent NIH 3T3 cells by oncogenic c-Raf-1. Mol. Cell. Biol. 17, 2576–2586 ( 1997).
Sewing, A., Wiseman, B., Lloyd, A. C. & Land, H. High-intensity Raf signal causes cell cycle arrest mediated by p21Cip1. Mol. Cell. Biol. 17, 5588–5597 ( 1997).
Lloyd, A. C. et al. Cooperating oncogenes converge to regulate cyclin/cdk complexes . Genes Dev 11, 663-677 (1997 ).
Cheng, M., Sexl, V., Sherr, C. J. & Roussel, M. F. Assembly of cyclin D-dependent kinase and titration of p27Kip1 regulated by mitogen-activated protein kinase kinase (MEK1). Proc. Natl Acad. Sci. USA 95, 1091–1096 ( 1998).
Aziz, N., Cherwinski, H. & McMahon, M. Complementation of defective colony-stimulating factor 1 receptor signaling and mitogenesis by Raf and v-Src. Mol. Cell. Biol. 19, 1101–1115 ( 1999).
Balmanno, K. & Cook, S. J. Sustained MAP kinase activation is required for the expression of cyclin D1, p21Cip1 and a subset of AP-1 proteins in CCL39 cells. Oncogene 18, 3085 –3097 (1999).
Peeper, D. S. et al. Ras signalling linked to the cell-cycle machinery by the retinoblastoma protein. Nature 386, 177– 181 (1997); erratum ibid. 386, 521 (1997).
Marshall, C. How do small GTPase signal transduction pathways regulate cell cycle entry? Curr. Opin. Cell Biol. 11, 732– 736 (1999).
Kerkhoff, E. & Rapp, U. R. Cell cycle targets of Ras/Raf signalling . Oncogene 17, 1457–1462 (1998).
Mateyak, M. K., Obaya, A. J. & Sedivy, J. M. c-Myc regulates cyclin D-Cdk4 and -Cdk6 activity but affects cell cycle progression at multiple independent points. Mol. Cell. Bio.l 19, 4672–4683 (1999).
Muller, D. et al. Cdk2-dependent phosphorylation of p27 facilitates its Myc-induced release from cyclin E/cdk2 complexes. Oncogene 15, 2561–2576 (1997).
Perez-Roger, I., Solomon, D. L., Sewing, A. & Land, H. Myc activation of cyclin E/Cdk2 kinase involves induction of cyclin E gene transcription and inhibition of p27(Kip1) binding to newly formed complexes. Oncogene 14, 2373–2381 ( 1997).
Vlach, J., Hennecke, S., Alevizopoulos, K., Conti, D. & Amati, B. Growth arrest by the cyclin-dependent kinase inhibitor p27Kip1 is abrogated by c-Myc. EMBO J. 15, 6595–6604 (1996).
Perez-Roger, I., Kim, S. H., Griffiths, B., Sewing, A. & Land, H. Cyclins D1 and D2 mediate Myc-induced proliferation via sequestration of p27(Kip1) and p21(Cip1). EMBO J. 18, 5310–5320 ( 1999).
Obaya, A. J., Mateyak, M. K. & Sedivy, J. M. Mysterious liaisons: the relationship between c-Myc and the cell cycle. Oncogene 18, 2934– 2941 (1999).
Dobrowolski, S., Harter, M. & Stacey, D. W. Cellular ras activity is required for passage through multiple points of the G0/G1 phase in BALB/c 3T3 cells. Mol. Cell. Biol. 14, 5441–5449 ( 1994).
Roche, S., Koegl, M. & Courtneidge, S. A. The phosphatidylinositol 3-kinase alpha is required for DNA synthesis induced by some, but not all, growth factors. Proc. Natl Acad. Sci. USA 91, 9185– 9189 (1994).
Bennett, A. M., Hausdorff, S. F., O'Reilly, A. M., Freeman, R. M. & Neel, B. G. Multiple requirements for SHPTP2 in epidermal growth factor-mediated cell cycle progression. Mol. Cell. Biol. 16, 1189–1202 (1996).
Takuwa, N. & Takuwa, Y. Ras activity late in G1 phase required for p27kip1 downregulation, passage through the restriction point, and entry into S phase in growth factor-stimulated NIH 3T3 fibroblasts . Mol. Cell. Biol. 17, 5348– 5358 (1997).
Jones, S. M., Klinghoffer, R., Prestwich, G. D., Toker, A. & Kazlauskas, A. PDGF induces an early and a late wave of PI 3-kinase activity, and only the late wave is required for progression through G1. Curr. Biol. 9, 512– 521 (1999).
Balciunaite, E., Jones, S. M., Toker, A. & Kazlauskas, A. PDGF initiates two distinct phases of PKC activity that make unequal contributions to the G0 to S transition. Curr. Biol. 10, 261– 267 (2000).
Alberts, B. et al. Molecular Biology of The Cell (eds Robetson, M. & Adams, R.) (Garland, New York, 1994).
Sherr, C. J. G1 phase progression: cycling on cue. Cell 79, 551–555 (1994).
Sherr, C. J. & Roberts, J. M. CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev. 13, 1501–1512 (1999).
Land, H., Parada, L. F. & Weinberg, R. A. Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature 304, 596–602 (1983).
Land, H., Chen, A. C., Morgenstern, J. P., Parada, L. F. & Weinberg, R. A. Behavior of myc and ras oncogenes in transformation of rat embryo fibroblasts. Mol. Cell. Biol. 6, 1917–1925 (1986).
Leone, G., DeGregori, J., Sears, R., Jakoi, L. & Nevins, J. R. Myc and Ras collaborate in inducing accumulation of active cyclin E/Cdk2 and E2F. Nature 387, 422–426 (1997); erratum ibid. 387, 932 (1997).
Amati, B., Alevizopoulos, K. & Vlach, J. Myc and the cell cycle. Front. Biosci. 3, D250–D268 (1998).
Daub, H., Weiss, F. U., Wallasch, C. & Ullrich, A. Role of transactivation of the EGF receptor in signalling by G-protein-coupled receptors. Nature 379, 557– 560 (1996).
Prenzel, N. et al. EGF receptor transactivation by G-protein-coupled receptors requires metalloproteinase cleavage of proHB-EGF. Nature 402, 884–888 (1999).
Valius, M. & Kazlauskas, A. Phospholipase C-gamma 1 and phosphatidylinositol 3 kinase are the downstream mediators of the PDGF receptor's mitogenic signal . Cell 73, 321–334 (1993).
Klinghoffer, R. A., Duckworth, B., Valius, M., Cantley, L. & Kazlauskas, A. Platelet-derived growth factor-dependent activation of phosphatidylinositol 3-kinase is regulated by receptor binding of SH2-domain-containing proteins which influence Ras activity. Mol. Cell. Biol. 16, 5905–5914 (1996).
Roussel, M. F., Theodoras, A. M., Pagano, M. & Sherr, C. J. Rescue of defective mitogenic signaling by D-type cyclins. Proc. Natl Acad. Sci. USA 92, 6837–6841 (1995).
Acknowledgements
We would like to thank M. Roussel for both the NIH3T3 cells expressing inducible MEK1* and the c-Myc retrovirus. We also appreciate the critical input of C. Sherr, M. Roussel, J. Pledger, C. Stiles, T. Langan, E. Balciunaite, M. Nickas and Y. Ikuno. This work was supported by an NIH grant (to A.K.) and an NIH postdoctoral fellowship (to S.M.J).
Author information
Authors and Affiliations
Corresponding author
Supplementary information
Figure 1 Continuous exposure to PDGF for less than 10 h compromises the mitogenic response.
Figure 2 MEK activity is necessary for the initial phase of signalling. (PDF 122 kb)
Figure 3 Expression of c-Myc to physiological levels does not deregulate growth factor-dependence for cell-cycle progression.
Figure 4 A 30-min pulse of PDGF is required to fully activate Erk and elevate c-Myc.
Figure 5 The second phase of signalling is transient and can be driven by synthetic PI(3)K lipid products.
Rights and permissions
About this article
Cite this article
Jones, S., Kazlauskas, A. Growth-factor-dependent mitogenesis requires two distinct phases of signalling . Nat Cell Biol 3, 165–172 (2001). https://doi.org/10.1038/35055073
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/35055073
This article is cited by
-
Neurodevelopmental disorders, like cancer, are connected to impaired chromatin remodelers, PI3K/mTOR, and PAK1-regulated MAPK
Biophysical Reviews (2023)
-
Targeting CDK4 and CDK6 in cancer
Nature Reviews Cancer (2022)
-
Relating individual cell division events to single-cell ERK and Akt activity time courses
Scientific Reports (2022)
-
A review of the biological and clinical implications of RAS-MAPK pathway alterations in neuroblastoma
Journal of Experimental & Clinical Cancer Research (2021)
-
Metabolic landscapes in sarcomas
Journal of Hematology & Oncology (2021)