Biochemical and molecular analysis of the interaction between ERK2 MAP kinase and hypoxia inducible factor-1α

Athanasios Karapetsas, Antonis Giannakakis, Maria Pavlaki, Mihalis Panayiotidis, Raphael Sandaltzopoulos, Alex Galanis

    Research output: Contribution to journalArticle

    Abstract

    The mitogen activated protein kinase (MAPK) signaling pathways play significant roles in fundamental cellular processes, such as cell growth and differentiation. It has been shown that the specificity and efficacy of phosphorylation by MAP kinases rely upon distinct MAPK-docking domains (D-domains) found in a wide range of MAPK substrates including the ETS-transcription factor Elk-1. Importantly, the MAPK signaling cascade converges with the hypoxia-induced signaling pathway. The key regulator of hypoxia signaling is the heterodimeric transcription factor hypoxia inducible factor-1 (HIF-1). The a-subunit of HIF-1 (HIF-1a) is a substrate for the ERK2 MAP kinase. Unraveling the interplay of these main signaling systems is a prerequisite for understanding their role in tumor growth, a situation sustained by simultaneous mitogenic and hypoxic signals. In this work, we investigated the molecular cues that direct HIF-1a recognition and phosphorylation by ERK2. We showed that HIF-1a possesses a MAPK docking domain. Utilizing surface plasmon resonance (SPR) methodologies we demonstrated efficient binding between HIF-1a and ERK2, with a K(D) value in the low micromolar range. Although, the D-domain did not contribute to the above interaction significantly, it could act in trans by recruiting ERK2 and conferring responsiveness to poor ERK substrates. These results indicate that, via its conserved D-domain, HIF-1a could serve as a platform for ERK2 in the nucleus of the cell, thus potentially facilitating phosphorylation of other ERK2 substrates. The identification of an ERK2 recognition domain on HIF-1a opens new avenues for the analysis of HIF-1a-related ERK2 signaling and may allow designing of interfering compounds.
    Original languageEnglish
    Pages (from-to)1582-1590
    Number of pages9
    JournalInternational Journal of Biochemistry and Cell Biology
    Volume43
    Issue number11
    DOIs
    Publication statusPublished - Nov 2011

    Fingerprint

    Hypoxia-Inducible Factor 1
    Mitogen-Activated Protein Kinases
    Phosphotransferases
    Phosphorylation
    ets-Domain Protein Elk-1
    Surface Plasmon Resonance
    Growth
    Cell Nucleus
    Cues
    Cell Differentiation
    Transcription Factors
    Neoplasms

    Cite this

    Karapetsas, Athanasios ; Giannakakis, Antonis ; Pavlaki, Maria ; Panayiotidis, Mihalis ; Sandaltzopoulos, Raphael ; Galanis, Alex. / Biochemical and molecular analysis of the interaction between ERK2 MAP kinase and hypoxia inducible factor-1α. In: International Journal of Biochemistry and Cell Biology. 2011 ; Vol. 43, No. 11. pp. 1582-1590.
    @article{5f22c837be844635b724bbe1650546d4,
    title = "Biochemical and molecular analysis of the interaction between ERK2 MAP kinase and hypoxia inducible factor-1α",
    abstract = "The mitogen activated protein kinase (MAPK) signaling pathways play significant roles in fundamental cellular processes, such as cell growth and differentiation. It has been shown that the specificity and efficacy of phosphorylation by MAP kinases rely upon distinct MAPK-docking domains (D-domains) found in a wide range of MAPK substrates including the ETS-transcription factor Elk-1. Importantly, the MAPK signaling cascade converges with the hypoxia-induced signaling pathway. The key regulator of hypoxia signaling is the heterodimeric transcription factor hypoxia inducible factor-1 (HIF-1). The a-subunit of HIF-1 (HIF-1a) is a substrate for the ERK2 MAP kinase. Unraveling the interplay of these main signaling systems is a prerequisite for understanding their role in tumor growth, a situation sustained by simultaneous mitogenic and hypoxic signals. In this work, we investigated the molecular cues that direct HIF-1a recognition and phosphorylation by ERK2. We showed that HIF-1a possesses a MAPK docking domain. Utilizing surface plasmon resonance (SPR) methodologies we demonstrated efficient binding between HIF-1a and ERK2, with a K(D) value in the low micromolar range. Although, the D-domain did not contribute to the above interaction significantly, it could act in trans by recruiting ERK2 and conferring responsiveness to poor ERK substrates. These results indicate that, via its conserved D-domain, HIF-1a could serve as a platform for ERK2 in the nucleus of the cell, thus potentially facilitating phosphorylation of other ERK2 substrates. The identification of an ERK2 recognition domain on HIF-1a opens new avenues for the analysis of HIF-1a-related ERK2 signaling and may allow designing of interfering compounds.",
    author = "Athanasios Karapetsas and Antonis Giannakakis and Maria Pavlaki and Mihalis Panayiotidis and Raphael Sandaltzopoulos and Alex Galanis",
    note = "Copyright {\circledC} 2011 Elsevier Ltd. All rights reserved.",
    year = "2011",
    month = "11",
    doi = "10.1016/j.biocel.2011.07.007",
    language = "English",
    volume = "43",
    pages = "1582--1590",
    journal = "International Journal of Biochemistry and Cell Biology",
    issn = "1357-2725",
    publisher = "Elsevier Limited",
    number = "11",

    }

    Biochemical and molecular analysis of the interaction between ERK2 MAP kinase and hypoxia inducible factor-1α. / Karapetsas, Athanasios; Giannakakis, Antonis; Pavlaki, Maria; Panayiotidis, Mihalis; Sandaltzopoulos, Raphael; Galanis, Alex.

    In: International Journal of Biochemistry and Cell Biology, Vol. 43, No. 11, 11.2011, p. 1582-1590.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Biochemical and molecular analysis of the interaction between ERK2 MAP kinase and hypoxia inducible factor-1α

    AU - Karapetsas, Athanasios

    AU - Giannakakis, Antonis

    AU - Pavlaki, Maria

    AU - Panayiotidis, Mihalis

    AU - Sandaltzopoulos, Raphael

    AU - Galanis, Alex

    N1 - Copyright © 2011 Elsevier Ltd. All rights reserved.

    PY - 2011/11

    Y1 - 2011/11

    N2 - The mitogen activated protein kinase (MAPK) signaling pathways play significant roles in fundamental cellular processes, such as cell growth and differentiation. It has been shown that the specificity and efficacy of phosphorylation by MAP kinases rely upon distinct MAPK-docking domains (D-domains) found in a wide range of MAPK substrates including the ETS-transcription factor Elk-1. Importantly, the MAPK signaling cascade converges with the hypoxia-induced signaling pathway. The key regulator of hypoxia signaling is the heterodimeric transcription factor hypoxia inducible factor-1 (HIF-1). The a-subunit of HIF-1 (HIF-1a) is a substrate for the ERK2 MAP kinase. Unraveling the interplay of these main signaling systems is a prerequisite for understanding their role in tumor growth, a situation sustained by simultaneous mitogenic and hypoxic signals. In this work, we investigated the molecular cues that direct HIF-1a recognition and phosphorylation by ERK2. We showed that HIF-1a possesses a MAPK docking domain. Utilizing surface plasmon resonance (SPR) methodologies we demonstrated efficient binding between HIF-1a and ERK2, with a K(D) value in the low micromolar range. Although, the D-domain did not contribute to the above interaction significantly, it could act in trans by recruiting ERK2 and conferring responsiveness to poor ERK substrates. These results indicate that, via its conserved D-domain, HIF-1a could serve as a platform for ERK2 in the nucleus of the cell, thus potentially facilitating phosphorylation of other ERK2 substrates. The identification of an ERK2 recognition domain on HIF-1a opens new avenues for the analysis of HIF-1a-related ERK2 signaling and may allow designing of interfering compounds.

    AB - The mitogen activated protein kinase (MAPK) signaling pathways play significant roles in fundamental cellular processes, such as cell growth and differentiation. It has been shown that the specificity and efficacy of phosphorylation by MAP kinases rely upon distinct MAPK-docking domains (D-domains) found in a wide range of MAPK substrates including the ETS-transcription factor Elk-1. Importantly, the MAPK signaling cascade converges with the hypoxia-induced signaling pathway. The key regulator of hypoxia signaling is the heterodimeric transcription factor hypoxia inducible factor-1 (HIF-1). The a-subunit of HIF-1 (HIF-1a) is a substrate for the ERK2 MAP kinase. Unraveling the interplay of these main signaling systems is a prerequisite for understanding their role in tumor growth, a situation sustained by simultaneous mitogenic and hypoxic signals. In this work, we investigated the molecular cues that direct HIF-1a recognition and phosphorylation by ERK2. We showed that HIF-1a possesses a MAPK docking domain. Utilizing surface plasmon resonance (SPR) methodologies we demonstrated efficient binding between HIF-1a and ERK2, with a K(D) value in the low micromolar range. Although, the D-domain did not contribute to the above interaction significantly, it could act in trans by recruiting ERK2 and conferring responsiveness to poor ERK substrates. These results indicate that, via its conserved D-domain, HIF-1a could serve as a platform for ERK2 in the nucleus of the cell, thus potentially facilitating phosphorylation of other ERK2 substrates. The identification of an ERK2 recognition domain on HIF-1a opens new avenues for the analysis of HIF-1a-related ERK2 signaling and may allow designing of interfering compounds.

    U2 - 10.1016/j.biocel.2011.07.007

    DO - 10.1016/j.biocel.2011.07.007

    M3 - Article

    VL - 43

    SP - 1582

    EP - 1590

    JO - International Journal of Biochemistry and Cell Biology

    JF - International Journal of Biochemistry and Cell Biology

    SN - 1357-2725

    IS - 11

    ER -