Selective SUMO modification of cAMP-specific phosphodiesterase-4D5 (PDE4D5) regulates the functional consequences of phosphorylation by PKA and ERK

Xiang Li, Suryakiran Vadrevu, Allan Dunlop, Jon Day, Noopur Advant, Jessica Troeger, Enno Klussmann, Ellis Jaffray, Ron T. Hay, David R. Adams, Miles D. Houslay, George S. Baillie

    Research output: Contribution to journalArticle

    27 Citations (Scopus)

    Abstract

    Enzymes from the PDE (phosphodiesterase) 4 cAMP-specific PDE family are crucial for the maintenance of compartmentalized cAMP responses in many cell types. Regulation of PDE activity can be achieved via post-translational modification such as phosphorylation by ERK (extracellular-signal-regulated kinase) MAPKs (mitogen-activated protein kinases) and PKA (protein kinase A). In the present paper, we report for the first time that PDE4 isoforms from the PDE4A and PDE4D subfamilies can be selectively modified by SUMO (small ubiquitin-related modifier). We have identified a single SUMO site within a consensus tetrapeptide motif, psi KXE (where psi represents a hydrophobic residue), which lies in the catalytic unit of these enzymes. SUMO modification of PDE4 at this site was observed upon overexpression of the SUMO E3 ligase PIASy [protein inhibitor of activated STAT (signal transducer and activator of transcription) Y] in HEK (human embryonic kidney)-293 cells and we identify PIASy as a novel binding partner for long PDE4 isoforms. Site-directed mutagenesis of the acceptor lysine residue ablated conjugation of PDE4 with SUMO, suggesting the presence of a single SUMO site in the first subdomain of the conserved PDE4 catalytic unit This observation was supported by both cell-free in vitro SUMOylation assays and analysis of SUMOylated spot-immobilized peptide arrays. SUMO modification of long PDE4 isoforms serves to augment their activation by PKA phosphorylation and repress their inhibition by ERK phosphorylation. Following ligation of beta-adrenergic receptors, SUMOylation of PDE4 isoforms sufficiently amplified PKA-stimulated PDE4 activity to reduce markedly the PKA phosphorylation status of the beta(2)-adrenergic receptor. These results highlight a new means whereby cells might achieve the selective regulation of the activity of cAMP-specific PDE4 enyzmes.

    Original languageEnglish
    Pages (from-to)55-65
    Number of pages11
    JournalBiochemical Journal
    Volume428
    DOIs
    Publication statusPublished - 15 May 2010

    Keywords

    • cAMP
    • peptide array
    • phosphodiesterase 4 (PDE4)
    • phosphorylation
    • protein inhibitor of activated signal transducer and activator of transcription Y (PIASy)
    • small ubiquitin-related modified (SUMO)
    • PROTEIN-KINASE-A
    • CYCLIC-AMP PHOSPHODIESTERASE
    • N-TERMINAL REGION
    • BETA-ARRESTIN
    • MEDIATED PHOSPHORYLATION
    • MEMBRANE ASSOCIATION
    • MAP KINASE
    • CROSS-TALK
    • IN-VIVO
    • ACTIVATION

    Cite this

    Li, Xiang ; Vadrevu, Suryakiran ; Dunlop, Allan ; Day, Jon ; Advant, Noopur ; Troeger, Jessica ; Klussmann, Enno ; Jaffray, Ellis ; Hay, Ron T. ; Adams, David R. ; Houslay, Miles D. ; Baillie, George S. / Selective SUMO modification of cAMP-specific phosphodiesterase-4D5 (PDE4D5) regulates the functional consequences of phosphorylation by PKA and ERK. In: Biochemical Journal. 2010 ; Vol. 428. pp. 55-65.
    @article{863a48eeec0c4837b7e3dbb50b385963,
    title = "Selective SUMO modification of cAMP-specific phosphodiesterase-4D5 (PDE4D5) regulates the functional consequences of phosphorylation by PKA and ERK",
    abstract = "Enzymes from the PDE (phosphodiesterase) 4 cAMP-specific PDE family are crucial for the maintenance of compartmentalized cAMP responses in many cell types. Regulation of PDE activity can be achieved via post-translational modification such as phosphorylation by ERK (extracellular-signal-regulated kinase) MAPKs (mitogen-activated protein kinases) and PKA (protein kinase A). In the present paper, we report for the first time that PDE4 isoforms from the PDE4A and PDE4D subfamilies can be selectively modified by SUMO (small ubiquitin-related modifier). We have identified a single SUMO site within a consensus tetrapeptide motif, psi KXE (where psi represents a hydrophobic residue), which lies in the catalytic unit of these enzymes. SUMO modification of PDE4 at this site was observed upon overexpression of the SUMO E3 ligase PIASy [protein inhibitor of activated STAT (signal transducer and activator of transcription) Y] in HEK (human embryonic kidney)-293 cells and we identify PIASy as a novel binding partner for long PDE4 isoforms. Site-directed mutagenesis of the acceptor lysine residue ablated conjugation of PDE4 with SUMO, suggesting the presence of a single SUMO site in the first subdomain of the conserved PDE4 catalytic unit This observation was supported by both cell-free in vitro SUMOylation assays and analysis of SUMOylated spot-immobilized peptide arrays. SUMO modification of long PDE4 isoforms serves to augment their activation by PKA phosphorylation and repress their inhibition by ERK phosphorylation. Following ligation of beta-adrenergic receptors, SUMOylation of PDE4 isoforms sufficiently amplified PKA-stimulated PDE4 activity to reduce markedly the PKA phosphorylation status of the beta(2)-adrenergic receptor. These results highlight a new means whereby cells might achieve the selective regulation of the activity of cAMP-specific PDE4 enyzmes.",
    keywords = "cAMP, peptide array, phosphodiesterase 4 (PDE4), phosphorylation, protein inhibitor of activated signal transducer and activator of transcription Y (PIASy), small ubiquitin-related modified (SUMO), PROTEIN-KINASE-A, CYCLIC-AMP PHOSPHODIESTERASE, N-TERMINAL REGION, BETA-ARRESTIN, MEDIATED PHOSPHORYLATION, MEMBRANE ASSOCIATION, MAP KINASE, CROSS-TALK, IN-VIVO, ACTIVATION",
    author = "Xiang Li and Suryakiran Vadrevu and Allan Dunlop and Jon Day and Noopur Advant and Jessica Troeger and Enno Klussmann and Ellis Jaffray and Hay, {Ron T.} and Adams, {David R.} and Houslay, {Miles D.} and Baillie, {George S.}",
    year = "2010",
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    doi = "10.1042/BJ20091672",
    language = "English",
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    Li, X, Vadrevu, S, Dunlop, A, Day, J, Advant, N, Troeger, J, Klussmann, E, Jaffray, E, Hay, RT, Adams, DR, Houslay, MD & Baillie, GS 2010, 'Selective SUMO modification of cAMP-specific phosphodiesterase-4D5 (PDE4D5) regulates the functional consequences of phosphorylation by PKA and ERK', Biochemical Journal, vol. 428, pp. 55-65. https://doi.org/10.1042/BJ20091672

    Selective SUMO modification of cAMP-specific phosphodiesterase-4D5 (PDE4D5) regulates the functional consequences of phosphorylation by PKA and ERK. / Li, Xiang; Vadrevu, Suryakiran; Dunlop, Allan; Day, Jon; Advant, Noopur; Troeger, Jessica; Klussmann, Enno; Jaffray, Ellis; Hay, Ron T.; Adams, David R.; Houslay, Miles D.; Baillie, George S.

    In: Biochemical Journal, Vol. 428, 15.05.2010, p. 55-65.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Selective SUMO modification of cAMP-specific phosphodiesterase-4D5 (PDE4D5) regulates the functional consequences of phosphorylation by PKA and ERK

    AU - Li, Xiang

    AU - Vadrevu, Suryakiran

    AU - Dunlop, Allan

    AU - Day, Jon

    AU - Advant, Noopur

    AU - Troeger, Jessica

    AU - Klussmann, Enno

    AU - Jaffray, Ellis

    AU - Hay, Ron T.

    AU - Adams, David R.

    AU - Houslay, Miles D.

    AU - Baillie, George S.

    PY - 2010/5/15

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    N2 - Enzymes from the PDE (phosphodiesterase) 4 cAMP-specific PDE family are crucial for the maintenance of compartmentalized cAMP responses in many cell types. Regulation of PDE activity can be achieved via post-translational modification such as phosphorylation by ERK (extracellular-signal-regulated kinase) MAPKs (mitogen-activated protein kinases) and PKA (protein kinase A). In the present paper, we report for the first time that PDE4 isoforms from the PDE4A and PDE4D subfamilies can be selectively modified by SUMO (small ubiquitin-related modifier). We have identified a single SUMO site within a consensus tetrapeptide motif, psi KXE (where psi represents a hydrophobic residue), which lies in the catalytic unit of these enzymes. SUMO modification of PDE4 at this site was observed upon overexpression of the SUMO E3 ligase PIASy [protein inhibitor of activated STAT (signal transducer and activator of transcription) Y] in HEK (human embryonic kidney)-293 cells and we identify PIASy as a novel binding partner for long PDE4 isoforms. Site-directed mutagenesis of the acceptor lysine residue ablated conjugation of PDE4 with SUMO, suggesting the presence of a single SUMO site in the first subdomain of the conserved PDE4 catalytic unit This observation was supported by both cell-free in vitro SUMOylation assays and analysis of SUMOylated spot-immobilized peptide arrays. SUMO modification of long PDE4 isoforms serves to augment their activation by PKA phosphorylation and repress their inhibition by ERK phosphorylation. Following ligation of beta-adrenergic receptors, SUMOylation of PDE4 isoforms sufficiently amplified PKA-stimulated PDE4 activity to reduce markedly the PKA phosphorylation status of the beta(2)-adrenergic receptor. These results highlight a new means whereby cells might achieve the selective regulation of the activity of cAMP-specific PDE4 enyzmes.

    AB - Enzymes from the PDE (phosphodiesterase) 4 cAMP-specific PDE family are crucial for the maintenance of compartmentalized cAMP responses in many cell types. Regulation of PDE activity can be achieved via post-translational modification such as phosphorylation by ERK (extracellular-signal-regulated kinase) MAPKs (mitogen-activated protein kinases) and PKA (protein kinase A). In the present paper, we report for the first time that PDE4 isoforms from the PDE4A and PDE4D subfamilies can be selectively modified by SUMO (small ubiquitin-related modifier). We have identified a single SUMO site within a consensus tetrapeptide motif, psi KXE (where psi represents a hydrophobic residue), which lies in the catalytic unit of these enzymes. SUMO modification of PDE4 at this site was observed upon overexpression of the SUMO E3 ligase PIASy [protein inhibitor of activated STAT (signal transducer and activator of transcription) Y] in HEK (human embryonic kidney)-293 cells and we identify PIASy as a novel binding partner for long PDE4 isoforms. Site-directed mutagenesis of the acceptor lysine residue ablated conjugation of PDE4 with SUMO, suggesting the presence of a single SUMO site in the first subdomain of the conserved PDE4 catalytic unit This observation was supported by both cell-free in vitro SUMOylation assays and analysis of SUMOylated spot-immobilized peptide arrays. SUMO modification of long PDE4 isoforms serves to augment their activation by PKA phosphorylation and repress their inhibition by ERK phosphorylation. Following ligation of beta-adrenergic receptors, SUMOylation of PDE4 isoforms sufficiently amplified PKA-stimulated PDE4 activity to reduce markedly the PKA phosphorylation status of the beta(2)-adrenergic receptor. These results highlight a new means whereby cells might achieve the selective regulation of the activity of cAMP-specific PDE4 enyzmes.

    KW - cAMP

    KW - peptide array

    KW - phosphodiesterase 4 (PDE4)

    KW - phosphorylation

    KW - protein inhibitor of activated signal transducer and activator of transcription Y (PIASy)

    KW - small ubiquitin-related modified (SUMO)

    KW - PROTEIN-KINASE-A

    KW - CYCLIC-AMP PHOSPHODIESTERASE

    KW - N-TERMINAL REGION

    KW - BETA-ARRESTIN

    KW - MEDIATED PHOSPHORYLATION

    KW - MEMBRANE ASSOCIATION

    KW - MAP KINASE

    KW - CROSS-TALK

    KW - IN-VIVO

    KW - ACTIVATION

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    DO - 10.1042/BJ20091672

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    VL - 428

    SP - 55

    EP - 65

    JO - Biochemical Journal

    JF - Biochemical Journal

    SN - 0264-6021

    ER -