AKAP79 inhibits calcineurin through a site distinct from the immunophilin-binding region.
Targeting of protein kinases and phosphatases provides additional specificity to substrate selectivity in cellular signaling. In the case of the Ca2+/calmodulin-dependent protein phosphatase calcineurin, AKAP79 has been shown to bind calcineurin and inhibit its activity in vitro (Coghlan, V., Perrino, B. A., Howard, M., Langeberg, L. K., Hicks, J. B., Gallatin, W. M., and Scott, J. D. (1995) Science 267, 108-111). In the present study, we characterized the binding regions on calcineurin A (CnA) and AKAP79 that are important for this interaction. Residues 30-98 and 311-336 on CnA, and residues 108-280 on AKAP79 were found to be important for binding. The binding of CnA by AKAP79 does not require the calcineurin B subunit, and occurs in a region distinct from where the immunosuppressant-immunophilin complex bind. AKAP79 also bound to CnA in cells transfected with AKAP79 and CnA. To determine the function of AKAP79-calcineurin interaction in intact cells, we measured the dephosphorylation and subsequent activation of NFAT, a transcription factor that is a substrate for calcineurin. Overexpression of AKAP79 inhibited NFAT dephosphorylation, resulting in a decrease in NFAT activation. These results demonstrated that AKAP79 can bind to and inhibit calcineurin activity in vivo, suggesting a physiological role for AKAP79-calcineurin interaction in NFAT-mediated signaling.
Kashishian
A
A
ICOS Corporation, Bothell, Washington 98021, USA.
Howard
M
M
Loh
C
C
Gallatin
W M
WM
Hoekstra
M F
MF
Lai
Y
Y
D001665:Binding Sites; D019703:Calcineurin / Q000378:metabolism; D065095:Calcineurin Inhibitors; D002352:Carrier Proteins; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000378:metabolism; D020104:Immunophilins / Q000378:metabolism; D008958:Models, Molecular; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D010446:Peptide Fragments / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000737:chemistry; D010750:Phosphoproteins / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism
D065095:Calcineurin Inhibitors; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D011506:Proteins; D014157:Transcription Factors; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases; D020104:Immunophilins