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Signaling diversity of PKA achieved via a Ca2+-cAMP-PKA oscillatory circuit

Abstract

Many protein kinases are key nodal signaling molecules that regulate a wide range of cellular functions. These functions may require complex spatiotemporal regulation of kinase activities. Here, we show that protein kinase A (PKA), Ca2+ and cyclic AMP (cAMP) oscillate in sync in insulin-secreting MIN6 beta cells, forming a highly integrated oscillatory circuit. We found that PKA activity was essential for this oscillatory circuit and was capable of not only initiating the signaling oscillations but also modulating their frequency, thereby diversifying the spatiotemporal control of downstream signaling. Our findings suggest that exquisite temporal control of kinase activity, mediated via signaling circuits resulting from cross-regulation of signaling pathways, can encode diverse inputs into temporal parameters such as oscillation frequency, which in turn contribute to proper regulation of complex cellular functions in a context-dependent manner.

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Figure 1: Oscillatory changes in PKA activity in single MIN6 beta cells.
Figure 2: Oscillatory changes in PKA activity, cAMP and Ca2+ dynamics are highly coordinated in MIN6 cells.
Figure 3: PKA activity is required for Ca2+ oscillation and tunes its frequency.
Figure 4: Direct activation of PKA triggers the oscillation of the circuit.
Figure 5: Oscillatory PKA activity confers spatial control of substrates.

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Acknowledgements

We thank J.-I. Miyazaki (Osaka University), G.G. Holz (SUNY Upstate Medical University) and G.H. Hart (Johns Hopkins University) for providing cell lines. We also thank X. Li (Johns Hopkins University) for initial technical assistance. This work was supported by US National Institutes of Health grants R01 DK073368 and DP1OD006419 (to J.Z.) and GM072024 and RR020839 (to A.L.).

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Contributions

Q.N. and J.Z. conceived the experimental aspect of the project and did the initial experiments; A.L. designed the modeling aspect of the project. Q.N. and N.-N.A.-H. performed the majority of the experiments. A.G. constructed the mathematical model and performed the simulations. X.G. performed the western analyses. M.D.A. designed and generated one of the biosensors. J.Z., Q.N. and A.L. wrote the manuscript.

Corresponding authors

Correspondence to Andre Levchenko or Jin Zhang.

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The authors declare no competing financial interests.

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Ni, Q., Ganesan, A., Aye-Han, NN. et al. Signaling diversity of PKA achieved via a Ca2+-cAMP-PKA oscillatory circuit. Nat Chem Biol 7, 34–40 (2011). https://doi.org/10.1038/nchembio.478

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