TY - JOUR
T1 - Regulation of nuclear PKA revealed by spatiotemporal manipulation of cyclic AMP
AU - Sample, Vedangi
AU - Dipilato, Lisa M.
AU - Yang, Jason H.
AU - Ni, Qiang
AU - Saucerman, Jeffrey J.
AU - Zhang, Jin
N1 - Funding Information:
We thank L. Levin (Weill Cornell Medical College, Cornell University) for the gift of sACt cDNA. We thank M. Houslay (Institute of Biomedical and Life Sciences, University of Glasgow) and K. Xiang (University of Illinois, Urbana-Champaign) for the gift of dnPDE4 isoforms. We thank L. Hersh (University of Kentucky College of Medicine) for giving us the A126.1B2 and A126.1B2 Catβ cell line. We also thank S. Mehta, G. Mo, T. Ueno, C. Pohlmeyer and T. Inoue for their technical help. This work was funded by US National Institutes of Health (NIH) grants R01 DK073368, DP1 OD006419 (to J.Z.), F31 DK074381 (to L.M.D.) and R01 HL094476, American Heart Association grant 0830470N (to J.J.S.) and NIH grant GM08715 (to J.H.Y.).
PY - 2012/4
Y1 - 2012/4
N2 - Understanding how specific cyclic AMP (cAMP) signals are organized and relayed to their effectors in different compartments of the cell to achieve functional specificity requires molecular tools that allow precise manipulation of cAMP in these compartments. Here we characterize a new method using bicarbonate-activatable and genetically targetable soluble adenylyl cyclase to control the location, kinetics and magnitude of the cAMP signal. Using this live-cell cAMP manipulation in conjunction with fluorescence imaging and mechanistic modeling, we uncovered the activation of a resident pool of protein kinase A (PKA) holoenzyme in the nuclei of HEK-293 cells, modifying the existing dogma of cAMP-PKA signaling in the nucleus. Furthermore, we show that phosphodiesterases and A-kinase anchoring proteins (AKAPs) are critical in shaping nuclear PKA responses. Collectively, our data suggest a new model in which AKAP-localized phosphodiesterases tune an activation threshold for nuclear PKA holoenzyme, thereby converting spatially distinct second messenger signals to temporally controlled nuclear kinase activity.
AB - Understanding how specific cyclic AMP (cAMP) signals are organized and relayed to their effectors in different compartments of the cell to achieve functional specificity requires molecular tools that allow precise manipulation of cAMP in these compartments. Here we characterize a new method using bicarbonate-activatable and genetically targetable soluble adenylyl cyclase to control the location, kinetics and magnitude of the cAMP signal. Using this live-cell cAMP manipulation in conjunction with fluorescence imaging and mechanistic modeling, we uncovered the activation of a resident pool of protein kinase A (PKA) holoenzyme in the nuclei of HEK-293 cells, modifying the existing dogma of cAMP-PKA signaling in the nucleus. Furthermore, we show that phosphodiesterases and A-kinase anchoring proteins (AKAPs) are critical in shaping nuclear PKA responses. Collectively, our data suggest a new model in which AKAP-localized phosphodiesterases tune an activation threshold for nuclear PKA holoenzyme, thereby converting spatially distinct second messenger signals to temporally controlled nuclear kinase activity.
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U2 - 10.1038/nchembio.799
DO - 10.1038/nchembio.799
M3 - Article
C2 - 22366721
AN - SCOPUS:84862806070
SN - 1552-4450
VL - 8
SP - 375
EP - 382
JO - Nature chemical biology
JF - Nature chemical biology
IS - 4
ER -