TY - JOUR
T1 - PKA and PDE4D3 anchoring to AKAP9 provides distinct regulation of cAMP signals at the centrosome
AU - Terrin, Anna
AU - Monterisi, Stefania
AU - Stangherlin, Alessandra
AU - Zoccarato, Anna
AU - Koschinski, Andreas
AU - Surdo, Nicoletta C.
AU - Mongillo, Marco
AU - Sawa, Akira
AU - Jordanides, Niove E.
AU - Mountford, Joanne C.
AU - Zaccolo, Manuela
PY - 2012/8
Y1 - 2012/8
N2 - Previous work has shown that the protein kinase A (PKA)-regulated phosphodiesterase (PDE) 4D3 binds to A kinase-anchoring proteins (AKAPs). One such protein, AKAP9, localizes to the centrosome. In this paper, we investigate whether a PKA-PDE4D3-AKAP9 complex can generate spatial compartmentalization of cyclic adenosine monophosphate (cAMP) signaling at the centrosome. Real-time imaging of fluorescence resonance energy transfer reporters shows that centrosomal PDE4D3 modulated a dynamic microdomain within which cAMP concentration selectively changed over the cell cycle. AP9-anchored, centrosomal PKA showed a reduced activation threshold as a consequence of increased auto-phosphorylation of its regulatory subunit at S114. Finally, disruption of the centrosomal cAMP microdomain by local displacement of PDE4D3 impaired cell cycle progression as a result of accumulation of cells in prophase. Our findings describe a novel mechanism of PKA activity regulation that relies on binding to AKAPs and consequent modulation of the enzyme activation threshold rather than on overall changes in cAMP levels. Further, we provide for the first time direct evidence that control of cell cycle progression relies on unique regulation of centrosomal cAMP/PKA signals.
AB - Previous work has shown that the protein kinase A (PKA)-regulated phosphodiesterase (PDE) 4D3 binds to A kinase-anchoring proteins (AKAPs). One such protein, AKAP9, localizes to the centrosome. In this paper, we investigate whether a PKA-PDE4D3-AKAP9 complex can generate spatial compartmentalization of cyclic adenosine monophosphate (cAMP) signaling at the centrosome. Real-time imaging of fluorescence resonance energy transfer reporters shows that centrosomal PDE4D3 modulated a dynamic microdomain within which cAMP concentration selectively changed over the cell cycle. AP9-anchored, centrosomal PKA showed a reduced activation threshold as a consequence of increased auto-phosphorylation of its regulatory subunit at S114. Finally, disruption of the centrosomal cAMP microdomain by local displacement of PDE4D3 impaired cell cycle progression as a result of accumulation of cells in prophase. Our findings describe a novel mechanism of PKA activity regulation that relies on binding to AKAPs and consequent modulation of the enzyme activation threshold rather than on overall changes in cAMP levels. Further, we provide for the first time direct evidence that control of cell cycle progression relies on unique regulation of centrosomal cAMP/PKA signals.
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U2 - 10.1083/jcb.201201059
DO - 10.1083/jcb.201201059
M3 - Article
C2 - 22908311
AN - SCOPUS:84866422570
SN - 0021-9525
VL - 198
SP - 607
EP - 621
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 4
ER -