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 Jaffrey, Ron T. Hay, David R. Adams, Miles D. Houslay, George S. Baillie

Research output: Contribution to journalArticle

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, ?KXE (where ? 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 ß-adrenergic receptors, SUMOylation of PDE4 isoforms sufficiently amplified PKA-stimulated PDE4 activity to reduce markedly the PKA phosphorylation status of the ß2-adrenergic receptor. These results highlight a new means whereby cells might achieve the selective regulation of the activity of cAMP-specific PDE4 enyzmes. © The Authors.

Original languageEnglish
Pages (from-to)55-65
Number of pages11
JournalBiochemical Journal
Volume428
Issue number1
DOIs
Publication statusPublished - 15 May 2010

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Extracellular Signal-Regulated MAP Kinases
Phosphoric Diester Hydrolases
Ubiquitin
Cyclic AMP-Dependent Protein Kinases
Phosphorylation
Protein Isoforms
Sumoylation
Adrenergic Receptors
Type 4 Cyclic Nucleotide Phosphodiesterase
Ubiquitin-Protein Ligases
Enzymes
Post Translational Protein Processing
Site-Directed Mutagenesis
Mitogen-Activated Protein Kinases
Transducers
Lysine
Ligation
Maintenance
Kidney
Peptides

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)

Cite this

Li, Xiang ; Vadrevu, Suryakiran ; Dunlop, Allan ; Day, Jon ; Advant, Noopur ; Troeger, Jessica ; Klussmann, Enno ; Jaffrey, 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, No. 1. pp. 55-65.
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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, ?KXE (where ? 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 {\ss}-adrenergic receptors, SUMOylation of PDE4 isoforms sufficiently amplified PKA-stimulated PDE4 activity to reduce markedly the PKA phosphorylation status of the {\ss}2-adrenergic receptor. These results highlight a new means whereby cells might achieve the selective regulation of the activity of cAMP-specific PDE4 enyzmes. {\circledC} The Authors.",
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Li, X, Vadrevu, S, Dunlop, A, Day, J, Advant, N, Troeger, J, Klussmann, E, Jaffrey, 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, no. 1, 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; Jaffrey, Ellis; Hay, Ron T.; Adams, David R.; Houslay, Miles D.; Baillie, George S.

In: Biochemical Journal, Vol. 428, No. 1, 15.05.2010, p. 55-65.

Research output: Contribution to journalArticle

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AU - Li, Xiang

AU - Vadrevu, Suryakiran

AU - Dunlop, Allan

AU - Day, Jon

AU - Advant, Noopur

AU - Troeger, Jessica

AU - Klussmann, Enno

AU - Jaffrey, Ellis

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AU - Houslay, Miles D.

AU - Baillie, George S.

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KW - Phosphorylation

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

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