Oligomerization conditions Mdm2-mediated efficient p53 polyubiquitylation but not its proteasomal degradation

Roland Hjerpe, Fabienne Aillet, Fernando Lopitz-Otsoa, Valerie Lang, Monica Torres-Ramos, Rosa Farras, Ronald T. Hay, Manuel S. Rodriguez

    Research output: Contribution to journalArticle

    9 Citations (Scopus)

    Abstract

    In normal cells p53 is maintained at low level through the action of the ubiquitin-proteasome system. As a consequence of p53 transcriptional activity, various regulators of this tumor suppressor are produced, forming a negative feedback loop tightly controlling p53 stability. One of the most prominent is the ubiquitin-ligase Mdm2. Here, we have used a transfer of signals strategy to study the p53 degradation process promoted by Mdm2 in the absence of p53 transcriptional activity. Our results show that in a p53 null background, transcriptionally silent p53-fusions require multiple N- and C-terminal signals to be optimally targeted to proteasomal degradation. As for WT p53, p53-fusions able to form tetramers are polyubiquitylated and optimally degraded by the proteasome. However, p53 molecules unable to oligomerize, show Mdm2-mediated polyubiquitylation deficiency but are still targeted to proteasome degradation in vitro and ex vivo. In the presence of Mdm2, nuclear shuttling of p53 monomeric fusions favours proteasome-dependent degradability but not its polyubiquitylation. In vitro, 265 proteasome fails to drive degradation of OD mutants in the presence of Mdm2, suggesting the contribution of additional cellular factors in this process. All together, our results indicate that Mdm2-mediated proteasome-dependent degradation of polyubiquitylation deficient p53 monomers is mechanistically possible, taking alternative pathways to better achieve their proteolysis. These results support the existence of additional levels to regulate p53 stability and activity acting on individual subunits of the functional tetramer. (C) 2010 Elsevier Ltd. All rights reserved.

    Original languageEnglish
    Pages (from-to)725-735
    Number of pages11
    JournalInternational Journal of Biochemistry & Cell Biology
    Volume42
    Issue number5
    DOIs
    Publication statusPublished - May 2010

    Keywords

    • p53
    • Polyubiquitylation
    • Degradation
    • Mdm2
    • Proteasome
    • TERMINAL DOMAIN
    • TRANSCRIPTIONAL ACTIVITY
    • PROTEIN-DEGRADATION
    • ACTIVATE P53
    • IN-VIVO
    • UBIQUITINATION
    • MDM2
    • MULTIPLE
    • IDENTIFICATION
    • MUTATIONS

    Cite this

    Hjerpe, Roland ; Aillet, Fabienne ; Lopitz-Otsoa, Fernando ; Lang, Valerie ; Torres-Ramos, Monica ; Farras, Rosa ; Hay, Ronald T. ; Rodriguez, Manuel S. / Oligomerization conditions Mdm2-mediated efficient p53 polyubiquitylation but not its proteasomal degradation. In: International Journal of Biochemistry & Cell Biology. 2010 ; Vol. 42, No. 5. pp. 725-735.
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    abstract = "In normal cells p53 is maintained at low level through the action of the ubiquitin-proteasome system. As a consequence of p53 transcriptional activity, various regulators of this tumor suppressor are produced, forming a negative feedback loop tightly controlling p53 stability. One of the most prominent is the ubiquitin-ligase Mdm2. Here, we have used a transfer of signals strategy to study the p53 degradation process promoted by Mdm2 in the absence of p53 transcriptional activity. Our results show that in a p53 null background, transcriptionally silent p53-fusions require multiple N- and C-terminal signals to be optimally targeted to proteasomal degradation. As for WT p53, p53-fusions able to form tetramers are polyubiquitylated and optimally degraded by the proteasome. However, p53 molecules unable to oligomerize, show Mdm2-mediated polyubiquitylation deficiency but are still targeted to proteasome degradation in vitro and ex vivo. In the presence of Mdm2, nuclear shuttling of p53 monomeric fusions favours proteasome-dependent degradability but not its polyubiquitylation. In vitro, 265 proteasome fails to drive degradation of OD mutants in the presence of Mdm2, suggesting the contribution of additional cellular factors in this process. All together, our results indicate that Mdm2-mediated proteasome-dependent degradation of polyubiquitylation deficient p53 monomers is mechanistically possible, taking alternative pathways to better achieve their proteolysis. These results support the existence of additional levels to regulate p53 stability and activity acting on individual subunits of the functional tetramer. (C) 2010 Elsevier Ltd. All rights reserved.",
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    Oligomerization conditions Mdm2-mediated efficient p53 polyubiquitylation but not its proteasomal degradation. / Hjerpe, Roland; Aillet, Fabienne; Lopitz-Otsoa, Fernando; Lang, Valerie; Torres-Ramos, Monica; Farras, Rosa; Hay, Ronald T.; Rodriguez, Manuel S.

    In: International Journal of Biochemistry & Cell Biology, Vol. 42, No. 5, 05.2010, p. 725-735.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Oligomerization conditions Mdm2-mediated efficient p53 polyubiquitylation but not its proteasomal degradation

    AU - Hjerpe, Roland

    AU - Aillet, Fabienne

    AU - Lopitz-Otsoa, Fernando

    AU - Lang, Valerie

    AU - Torres-Ramos, Monica

    AU - Farras, Rosa

    AU - Hay, Ronald T.

    AU - Rodriguez, Manuel S.

    PY - 2010/5

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    AB - In normal cells p53 is maintained at low level through the action of the ubiquitin-proteasome system. As a consequence of p53 transcriptional activity, various regulators of this tumor suppressor are produced, forming a negative feedback loop tightly controlling p53 stability. One of the most prominent is the ubiquitin-ligase Mdm2. Here, we have used a transfer of signals strategy to study the p53 degradation process promoted by Mdm2 in the absence of p53 transcriptional activity. Our results show that in a p53 null background, transcriptionally silent p53-fusions require multiple N- and C-terminal signals to be optimally targeted to proteasomal degradation. As for WT p53, p53-fusions able to form tetramers are polyubiquitylated and optimally degraded by the proteasome. However, p53 molecules unable to oligomerize, show Mdm2-mediated polyubiquitylation deficiency but are still targeted to proteasome degradation in vitro and ex vivo. In the presence of Mdm2, nuclear shuttling of p53 monomeric fusions favours proteasome-dependent degradability but not its polyubiquitylation. In vitro, 265 proteasome fails to drive degradation of OD mutants in the presence of Mdm2, suggesting the contribution of additional cellular factors in this process. All together, our results indicate that Mdm2-mediated proteasome-dependent degradation of polyubiquitylation deficient p53 monomers is mechanistically possible, taking alternative pathways to better achieve their proteolysis. These results support the existence of additional levels to regulate p53 stability and activity acting on individual subunits of the functional tetramer. (C) 2010 Elsevier Ltd. All rights reserved.

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    KW - Degradation

    KW - Mdm2

    KW - Proteasome

    KW - TERMINAL DOMAIN

    KW - TRANSCRIPTIONAL ACTIVITY

    KW - PROTEIN-DEGRADATION

    KW - ACTIVATE P53

    KW - IN-VIVO

    KW - UBIQUITINATION

    KW - MDM2

    KW - MULTIPLE

    KW - IDENTIFICATION

    KW - MUTATIONS

    U2 - 10.1016/j.biocel.2010.01.010

    DO - 10.1016/j.biocel.2010.01.010

    M3 - Article

    VL - 42

    SP - 725

    EP - 735

    JO - International Journal of Biochemistry & Cell Biology

    JF - International Journal of Biochemistry & Cell Biology

    SN - 1357-2725

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