Protein kinase A-dependent phosphorylation of Rap1 regulates its membrane localization and cell migration.
The small G protein Rap1 can mediate "inside-out signaling" by recruiting effectors to the plasma membrane that signal to pathways involved in cell adhesion and cell migration. This action relies on the membrane association of Rap1, which is dictated by post-translational prenylation as well as by a stretch of basic residues within its carboxyl terminus. One feature of this stretch of acidic residues is that it lies adjacent to a functional phosphorylation site for the cAMP-dependent protein kinase PKA. This phosphorylation has two effects on Rap1 action. One, it decreases the level of Rap1 activity as measured by GTP loading and the coupling of Rap1 to RapL, a Rap1 effector that couples Rap1 GTP loading to integrin activation. Two, it destabilizes the membrane localization of Rap1, promoting its translocation into the cytoplasm. These two actions, decreased GTP loading and decreased membrane localization, are related, as the translocation of Rap1-GTP into the cytoplasm is associated with its increased GTP hydrolysis and inactivation. The consequences of this phosphorylation in Rap1-dependent cell adhesion and cell migration were also examined. Active Rap1 mutants that lack this phosphorylation site had a minimal effect on cell adhesion but strongly reduced cell migration, when compared with an active Rap1 mutant that retained the phosphorylation site. This suggests that optimal cell migration is associated with cycles of Rap1 activation, membrane egress, and inactivation, and requires the regulated phosphorylation of Rap1 by PKA.
From the Vollum Institute, and.
Maho
M
Takahashi
Tara J
TJ
Dillon
Chang
C
Liu
Yumi
Y
Kariya
Zhiping
Z
Wang
Philip J S
PJ
Stork
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry*; D004789:Enzyme Activation; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D015398:Signal Transduction; D020827:rap1 GTP-Binding Proteins / Q000737:chemistry*
D017868:Cyclic AMP-Dependent Protein Kinases; D020827:rap1 GTP-Binding Proteins
Cell Adhesion; Cell Migration; Electrostatic Switch; Geranylgeranylation; Low Molecular Weight G Proteins; Phosphorylation; Protein Kinase A (PKA); Rap1; cAMP