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Analyzing Ras-Associated Cell Proliferation Signaling

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Cell Cycle Control

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1170))

Abstract

Ras-dependent signaling is an important regulator of cell cycle progression, proliferation, senescence, and apoptosis. Several of the downstream effectors of Ras play dual roles in each of these processes. Under one set of conditions, they promote cell cycle progression and proliferation; yet, in a different paradigm, they drive cell cycle arrest and apoptosis. Furthermore, there is cross talk between certain downstream effectors of Ras including the PI3K–AKT and Raf–MEK–ERK pathways. Here we describe a series of experiments used to dissect the effect of different Ras-dependent signaling pathways on cell cycle progression, proliferation, senescence, and apoptosis. Furthermore, we highlight the importance of consistent growth conditions of cells in culture when studying Ras-dependent signaling as we show that the activation of downstream effectors of Ras changes with the confluency at which the cells are grown.

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Acknowledgements

We would like to thank Channing J. Der for the kind gift of the GTPase-binding domain pGEX constructs used in the pulldown assays.

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Authors and Affiliations

  1. Department of Pharmacology and Physiology, Drexel University College of Medicine, 245 N. 15th Street, MS 488, Philadelphia, PA, 19102, USA

    Matthew C. Stout, Edgar Asiimwe, James R. Birkenstamm, Su Yeon Kim & Paul M. Campbell

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  1. Matthew C. Stout
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  2. Edgar Asiimwe
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  3. James R. Birkenstamm
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  4. Su Yeon Kim
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  5. Paul M. Campbell
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Corresponding author

Correspondence to Paul M. Campbell .

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Editors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, Pennsylvania, USA

    Eishi Noguchi

  2. Dept of Biochemistry & Molecular Biology, Drexel University College of Medicine, Philadelphia, USA

    Mariana C. Gadaleta

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Stout, M.C., Asiimwe, E., Birkenstamm, J.R., Kim, S.Y., Campbell, P.M. (2014). Analyzing Ras-Associated Cell Proliferation Signaling. In: Noguchi, E., Gadaleta, M. (eds) Cell Cycle Control. Methods in Molecular Biology, vol 1170. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0888-2_21

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  • DOI: https://doi.org/10.1007/978-1-4939-0888-2_21

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0887-5

  • Online ISBN: 978-1-4939-0888-2

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