Visualizing and quantitating the spatiotemporal regulation of Ras/ERK signaling by dual-specificity Mitogen-Activated Protein Phosphatases (MKPs)

Christopher J. Caunt, Andrew M. Kidger, Stephen M. Keyse (Lead / Corresponding author)

    Research output: Chapter in Book/Report/Conference proceedingChapter

    4 Citations (Scopus)

    Abstract

    The spatiotemporal regulation of the Ras/ERK pathway is critical in determining the physiological and pathophysiological outcome of signaling. Dual-specificity mitogen-activated protein kinase (MAPK) phos-phatases (DUSPs or MKPs) are key regulators of pathway activity and may also localize ERK to distinct subcellular locations. Here we present methods largely based on the use of high content microscopy to both visualize and quantitate the subcellular distribution of activated (p-ERK) and total ERK in popula-tions of mouse embryonic fibroblasts derived from mice lacking DUSP5, a nuclear ERK-specific MKP. Such methods in combination with rescue experiments using adenoviral vectors encoding wild-type and mutant forms of DUSP5 have allowed us to visualize specific defects in ERK regulation in these cells thus confirm-ing the role of this phosphatase as both a nuclear regulator of ERK activity and localization.
    Original languageEnglish
    Title of host publicationProtein tyrosine phosphatases
    Subtitle of host publicationmethods and protocols
    EditorsRafael Pulido
    Place of PublicationNew York
    PublisherSpringer
    Pages197-215
    Number of pages12
    ISBN (Electronic)9781493937462
    ISBN (Print)9781493937448
    DOIs
    Publication statusPublished - 2016

    Publication series

    NameMethods in molecular biology
    PublisherSpringer Science+Business Media
    Volume1447
    ISSN (Print)1064-3745

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  • Cite this

    Caunt, C. J., Kidger, A. M., & Keyse, S. M. (2016). Visualizing and quantitating the spatiotemporal regulation of Ras/ERK signaling by dual-specificity Mitogen-Activated Protein Phosphatases (MKPs). In R. Pulido (Ed.), Protein tyrosine phosphatases: methods and protocols (pp. 197-215). (Methods in molecular biology; Vol. 1447). Springer . https://doi.org/10.1007/978-1-4939-3746-2_12