The N terminus domain of type VI adenylyl cyclase mediates its inhibition by protein kinase C.
Previous results from our laboratory have shown that phosphorylation of type VI adenylyl cyclase (ACVI) by protein kinase C (PKC) caused suppression of adenylyl cyclase activity. In the present study, we investigated the role of the N terminus cytosolic domain of ACVI in this PKC-mediated inhibition of ACVI. Removal of amino acids 1 to 86 of ACVI or mutation of Ser(10) (a potential PKC phosphorylation site) into alanine significantly relieved the PKC-mediated inhibition and markedly reduced the PKC-evoked protein phosphorylation. PKC also effectively phosphorylated a recombinant N terminus cytosolic domain (amino acids 1-160) protein of ACVI and a synthetic peptide representing Ser(10). In addition, the amino acids 1 to 86 truncated mutant exhibited kinetic properties similar to those of the wild type. Taken together, these data demonstrate that the highly variable N terminus cytoplasmic domain of ACVI is a regulatory domain with a critical role in PKC-mediated suppression, which is a hallmark of this adenylyl cyclase isozyme. In addition, Ser(10) was found to serve as an acceptor for the PKC-mediated phosphorylating transfer of ACVI.
Lai
H L
HL
Division of Neuroscience, Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, Republic of China.
Lin
T H
TH
Kao
Y Y
YY
Lin
W J
WJ
Hwang
M J
MJ
Chern
Y
Y
D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000378:metabolism; D007313:Insecta; D009154:Mutation; D010766:Phosphorylation; D011487:Protein Conformation; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D012694:Serine / Q000235:genetics
D000067956:Adenylyl Cyclase Inhibitors; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D012694:Serine; D011493:Protein Kinase C; D000262:Adenylyl Cyclases; C121100:adenylyl cyclase 6