Abstract
A myriad of cellular processes instigated by growth factors are mediated by cell surface-associated receptor tyrosine kinases (RTKs). Subsequent downstream activation of signaling cascades, as well as their crosstalk, endows specificity in terms of the phenotypic outcome, e.g., cellular proliferation, migration, or differentiation. Such signaling diversity is exemplified by the ability of the epidermal growth factor receptor (EGFR) to stimulate different MAPK cascades, especially the ERK1/2 cascade. It has been shown that the ability of the ERK1/2 cascade to specify cell fate, such as cell migration, is dependent on signal duration governed by feedback control. Here we focus on one experimental system, MCF10A human mammary cells, and a phenotypic outcome of cell migration. We present methods to identify key components of underlying cascades and their effects on the migratory phenotype. We focus on profiling activation of signaling modules, as well as transcriptional regulation, emphasizing the high-throughput potential of such approaches.
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References
Hunter T (2000) Signaling–2000 and beyond. Cell 100(1):113–127.
Pawson T (2004) Specificity in signal transduction: from phosphotyrosine-SH2 domain interactions to complex cellular systems. Cell 116(2):191–203.
Citri A & Yarden Y (2006) EGF-ERBB signalling: towards the systems level. Nat Rev Mol Cell Biol 7(7):505–516.
Jones RB, Gordus A, Krall JA, & MacBeath G (2006) A quantitative protein interaction network for the ErbB receptors using protein microarrays. Nature 439(7073):168–174.
Katz M, Amit I, & Yarden Y (2007) Regulation of MAPKs by growth factors and receptor tyrosine kinases. Biochim Biophys Acta 1773(8):1161–1176.
Marshall CJ (1995) Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell 80(2):179–185.
Santos SD, Verveer PJ, & Bastiaens PI (2007) Growth factor-induced MAPK network topology shapes Erk response determining PC-12 cell fate. Nat Cell Biol 9(3):324–330.
Treisman R (1996) Regulation of transcription by MAP kinase cascades. Curr Opin Cell Biol 8(2):205–215.
Amit I, et al. (2007) A module of negative feedback regulators defines growth factor signaling. Nat Genet 39(4):503–512.
Perez OD & Nolan GP (2002) Simultaneous measurement of multiple active kinase states using polychromatic flow cytometry. Nat Biotechnol 20(2):155–162.
Krutzik PO, Irish JM, Nolan GP, & Perez OD (2004) Analysis of protein phosphorylation and cellular signaling events by flow cytometry: techniques and clinical applications. Clin Immunol 110(3):206–221.
Irish JM, et al. (2004) Single cell profiling of potentiated phospho-protein networks in cancer cells. Cell 118(2):217–228.
Sachs K, Perez O, Pe’er D, Lauffenburger DA, & Nolan GP (2005) Causal protein-signaling networks derived from multiparameter single-cell data. Science 308(5721):523–529.
Irie HY, et al. (2005) Distinct roles of Akt1 and Akt2 in regulating cell migration and epithelial-mesenchymal transition. J Cell Biol 171(6):1023–1034.
Acknowledgments
Our laboratory is supported by research grants from the National Cancer Institute (grant CA72981), the M.D. Moross Institute for Cancer Research and the Willner Family Center for Vascular Biology. Y.Y. is the incumbent of the Harold and Zelda Goldenberg Professorial Chair.
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Tarcic, G., Yarden, Y. (2010). MAP Kinase Activation by Receptor Tyrosine Kinases: In Control of Cell Migration. In: Seger, R. (eds) MAP Kinase Signaling Protocols. Methods in Molecular Biology, vol 661. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-795-2_7
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DOI: https://doi.org/10.1007/978-1-60761-795-2_7
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Publisher Name: Humana Press, Totowa, NJ
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