Spatial regulation of cyclic AMP-Epac1 signaling in cell adhesion by ERM proteins.
Epac1 is a guanine nucleotide exchange factor for the small G protein Rap and is involved in membrane-localized processes such as integrin-mediated cell adhesion and cell-cell junction formation. Cyclic AMP (cAMP) directly activates Epac1 by release of autoinhibition and in addition induces its translocation to the plasma membrane. Here, we show an additional mechanism of Epac1 recruitment, mediated by activated ezrin-radixin-moesin (ERM) proteins. Epac1 directly binds with its N-terminal 49 amino acids to ERM proteins in their open conformation. Receptor-induced activation of ERM proteins results in increased binding of Epac1 and consequently the clustered localization of Epac1 at the plasma membrane. Deletion of the N terminus of Epac1, as well as disruption of the Epac1-ERM interaction by an interfering radixin mutant or small interfering RNA (siRNA)-mediated depletion of the ERM proteins, impairs Epac1-mediated cell adhesion. We conclude that ERM proteins are involved in the spatial regulation of Epac1 and cooperate with cAMP- and Rap-mediated signaling to regulate adhesion to the extracellular matrix.
Gloerich
Martijn
M
Department of Physiological Chemistry, Center for Biomedical Genetics, and Cancer Genomics Center, University Medical Center Utrecht, Utrecht, Netherlands.
Ponsioen
Bas
B
Vliem
Marjolein J
MJ
Zhang
Zhongchun
Z
Zhao
Jun
J
Kooistra
Matthijs R
MR
Price
Leo S
LS
Ritsma
Laila
L
Zwartkruis
Fried J
FJ
Rehmann
Holger
H
Jalink
Kees
K
Bos
Johannes L
JL
D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D020798:Two-Hybrid System Techniques; D020827:rap1 GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D000099149:Ezrin
D000242:Cyclic AMP; D003598:Cytoskeletal Proteins; D020662:Guanine Nucleotide Exchange Factors; D008565:Membrane Proteins; D008840:Microfilament Proteins; D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins; D020827:rap1 GTP-Binding Proteins; D000099149:Ezrin; C116122:RAPGEF3 protein, human; C442864:RAPGEF4 protein, human; C071058:moesin; C071776:radixin