Adenylyl cyclases 5 and 6 underlie PIP3-dependent regulation
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Adenylyl cyclases 5 and 6 underlie PIP3-dependent regulation. / Reddy, Gopireddy Raghavender; Subramanian, Hariharan; Birk, Alexandra; Milde, Markus; Nikolaev, Viacheslav O; Bünemann, Moritz.
In: FASEB J, Vol. 29, No. 8, 08.2015, p. 3458-71.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Adenylyl cyclases 5 and 6 underlie PIP3-dependent regulation
AU - Reddy, Gopireddy Raghavender
AU - Subramanian, Hariharan
AU - Birk, Alexandra
AU - Milde, Markus
AU - Nikolaev, Viacheslav O
AU - Bünemann, Moritz
N1 - © FASEB.
PY - 2015/8
Y1 - 2015/8
N2 - Many different neurotransmitters and hormones control intracellular signaling by regulating the production of the second messenger cAMP. The function of the broadly expressed adenylyl cyclases (ACs) 5 and 6 is regulated by either stimulatory or inhibitory G proteins. By analyzing a well-known rebound stimulation phenomenon after withdrawal of Gi protein in atrial myocytes, we discovered that AC5 and -6 are tightly regulated by the second messenger PIP3. By monitoring cAMP levels in real time by means of Förster resonance energy transfer (FRET)-based biosensors, we reproduced the rebound stimulation in a heterologous expression system specifically for AC5 or -6. Strikingly, this cAMP rebound stimulation was completely blocked by the PI3K inhibitor wortmannin, both in atrial myocytes and in transfected human embryonic kidney cells. Similar effects were observed by heterologous expression of the PIP3 phosphatase and tensin homolog (PTEN). However, general kinase inhibitors or inhibitors of Akt had no effect, suggesting a PIP3-dependent mechanism. These findings demonstrate the existence of a novel general pathway for regulation of AC5 and -6 activity via PIP3 that leads to pronounced alterations of cytosolic cAMP levels.
AB - Many different neurotransmitters and hormones control intracellular signaling by regulating the production of the second messenger cAMP. The function of the broadly expressed adenylyl cyclases (ACs) 5 and 6 is regulated by either stimulatory or inhibitory G proteins. By analyzing a well-known rebound stimulation phenomenon after withdrawal of Gi protein in atrial myocytes, we discovered that AC5 and -6 are tightly regulated by the second messenger PIP3. By monitoring cAMP levels in real time by means of Förster resonance energy transfer (FRET)-based biosensors, we reproduced the rebound stimulation in a heterologous expression system specifically for AC5 or -6. Strikingly, this cAMP rebound stimulation was completely blocked by the PI3K inhibitor wortmannin, both in atrial myocytes and in transfected human embryonic kidney cells. Similar effects were observed by heterologous expression of the PIP3 phosphatase and tensin homolog (PTEN). However, general kinase inhibitors or inhibitors of Akt had no effect, suggesting a PIP3-dependent mechanism. These findings demonstrate the existence of a novel general pathway for regulation of AC5 and -6 activity via PIP3 that leads to pronounced alterations of cytosolic cAMP levels.
KW - Adenylyl Cyclases
KW - Cell Line
KW - Cell Line, Tumor
KW - Cyclic AMP
KW - GTP-Binding Protein alpha Subunits, Gi-Go
KW - HEK293 Cells
KW - HeLa Cells
KW - Humans
KW - PTEN Phosphohydrolase
KW - Phosphatidylinositol 3-Kinases
KW - Signal Transduction
U2 - 10.1096/fj.14-268466
DO - 10.1096/fj.14-268466
M3 - SCORING: Journal article
C2 - 25931510
VL - 29
SP - 3458
EP - 3471
JO - FASEB J
JF - FASEB J
SN - 0892-6638
IS - 8
ER -
