Cyclic Nucleotide-dependent Protein Kinases Target ARHGAP17 and ARHGEF6 Complexes in Platelets
|Title:||Cyclic Nucleotide-dependent Protein Kinases Target ARHGAP17 and ARHGEF6 Complexes in Platelets||Authors:||Nagy, Zoltan
Von Kriegsheim, Alex
Smolenski, Albert P.
|Permanent link:||http://hdl.handle.net/10197/7388||Date:||11-Dec-2015||Online since:||2016-10-27T01:00:09Z||Abstract:||Endothelial cells release prostacyclin (PGI2) and nitric oxide (NO) to inhibit platelet functions. PGI2 and NO effects are mediated by cyclic nucleotides, cAMP- and cGMP-dependent protein kinases (PKA, PKG), and largely unknown PKA and PKG substrate proteins. The small G-protein Rac1 plays a key role in platelets and was suggested to be a target of cyclic nucleotide signaling. We confirm that PKA and PKG activation reduces Rac1-GTP levels. Screening for potential mediators of this effect resulted in the identification of the Rac1-specific GTPase-activating protein ARHGAP17 and the guanine nucleotide exchange factor ARHGEF6 as new PKA and PKG substrates in platelets. We mapped the PKA/PKG phosphorylation sites to serine 702 on ARHGAP17 using Phos-tag gels and to serine 684 on ARHGEF6. We show that ARHGAP17 binds to the actin-regulating CIP4 protein in platelets and that Ser-702 phosphorylation interferes with this interaction. Reduced CIP4 binding results in enhanced inhibition of cell migration by ARHGAP17. Furthermore, we show that ARHGEF6 is constitutively linked to GIT1, a GAP of Arf family small G proteins, and that ARHGEF6 phosphorylation enables binding of the 14-3-3 adaptor protein to the ARHGEF6/GIT1 complex. PKA and PKG induced rearrangement of ARHGAP17- and ARHGEF6-associated protein complexes might contribute to Rac1 regulation and platelet inhibition.||Funding Details:||Irish Research Council
Science Foundation Ireland
|Type of material:||Journal Article||Publisher:||American Society for Biochemistry and Molecular Biology||Journal:||Journal of Biological Chemistry||Volume:||290||Issue:||50||Start page:||29974||End page:||29983||Copyright (published version):||2015 The American Society for Biochemistry and Molecular Biology, Inc.||Keywords:||Rac (Rac GTPase); Cyclic AMP (cAMP); cyclic GMP (cGMP); Phosphorylation; Platelet||DOI:||10.1074/jbc.M115.678003||Language:||en||Status of Item:||Peer reviewed|
|Appears in Collections:||Conway Institute Research Collection|
SBI Research Collection
Medicine Research Collection
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