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Control of PKA stability and signalling by the RING ligase praja2

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Abstract

Activation of G-protein-coupled receptors (GPCRs) mobilizes compartmentalized pulses of cyclic AMP. The main cellular effector of cAMP is protein kinase A (PKA), which is assembled as an inactive holoenzyme consisting of two regulatory (R) and two catalytic (PKAc) subunits. cAMP binding to R subunits dissociates the holoenzyme and releases the catalytic moiety, which phosphorylates a wide array of cellular proteins. Reassociation of PKAc and R components terminates the signal. Here we report that the RING ligase praja2 controls the stability of mammalian R subunits. Praja2 forms a stable complex with, and is phosphorylated by, PKA. Rising cAMP levels promote praja2-mediated ubiquitylation and subsequent proteolysis of compartmentalized R subunits, leading to sustained substrate phosphorylation by the activated kinase. Praja2 is required for efficient nuclear cAMP signalling and for PKA-mediated long-term memory. Thus, praja2 regulates the total concentration of R subunits, tuning the strength and duration of PKA signal output in response to cAMP.

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Figure 1: Praja2 binds to PKA R subunits.
Figure 2: Praja2 co-localizes with PKA R subunits.
Figure 3: Praja2 triggers degradation of R subunits.
Figure 4: Praja2 ubiquitylates RIIα/β subunits.
Figure 5: PKAc binds to and phosphorylates praja2.
Figure 6: Praja2 controls PKA signalling in cells.
Figure 7: Praja2 controls PKA signalling and LTP in rat brain.
Figure 8: Schematic representation of the role of praja2 in PKA signalling.

Change history

  • 22 March 2011

    In the version of this article initially published online Figures 2, 4 and 5 were mislabelled, and on page 5 line 21 a sentence has now been reworded for clarity.

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Acknowledgements

This work was supported by Grants from Associazione Italiana per la Ricerca sul Cancro (AIRC) and the Italian Ministry of University and Research (MIUR) 2008–2009 (2007KS47FW_002). E.S. was supported from Austrian Science Fund (FWF) Grant P22608 and Junior Researcher Support 2009 (University of Innsbruck). Special thanks to M. Gottesman, E. Avvedimento and K. Bister for discussion and critical reading of the manuscript, D. Viaggiano for help in immunohistochemistry and confocal microscopy analysis, R. D. Donne for help in carrying out experiments and S. McKnight and M. Ginsberg for providing the vectors for epitope-tagged R subunits. This manuscript is dedicated to the memory of M. Graziano.

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Authors

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L.L., L.A., E.S. and A.F. designed the experiments. L.L. carried out most of the experiments except for the following:

Figs 2c, 7c,d and Supplementary Figs S5 and S7b were carried out by A.S., C.S. and O.C.

Figs 1d and 5g were carried out by M.S., who also generated praja2-deletion mutants.

Figs 3f–h, 5c–f, i, j, m and Supplementary Fig. S6b were carried out by E.S.

Figs 2a,b, 5a, 6a and Supplementary Figs S3 and S4 were carried out by C.G.

Figs 6g,h, 7e and Supplementary Fig. S7a were carried out by A.C.

Fig. 7f,g were carried out by R.N.

L.L., E.S., L.A. and A.F. analysed the data.

A.F. wrote the manuscript with contributions from L.A. and E.S.

Corresponding author

Correspondence to Antonio Feliciello.

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The authors declare no competing financial interests.

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Lignitto, L., Carlucci, A., Sepe, M. et al. Control of PKA stability and signalling by the RING ligase praja2. Nat Cell Biol 13, 412–422 (2011). https://doi.org/10.1038/ncb2209

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