TY - JOUR
T1 - PIP3-Independent Activation of TorC2 and PKB at the Cell's Leading Edge Mediates Chemotaxis
AU - Kamimura, Yoichiro
AU - Xiong, Yuan
AU - Iglesias, Pablo A.
AU - Hoeller, Oliver
AU - Bolourani, Parvin
AU - Devreotes, Peter N.
N1 - Funding Information:
The authors wish to acknowledge members of the Devreotes lab, particularly Jonathan Franca-Koh and Stacey Willard, for critical reading. We thank Dr. R.R. Kay for pi3k1-5 − cells; Dr. M. Iijima for the PKBA disruption plasmid; Dr. K. Inoue for α-Talin B antibody; Dr. R.H. Insall for aleA − cells; Drs. R. Meile and R.A. Firtel for the PKBR1 disruption plasmid, PKBR1 cDNA, and helpful advice; Dr. G. Weeks for cAR1-expressing rasC − G − cells; and proteomics core facility in Johns Hopkins University, School of Medicine, for mass-spectrometry analysis. This work was supported by National Institutes of Health (NIH) GM 28007 and NIH GM 34933 to P.N.D., by Uehara memorial foundation to Y.K., and by GM 71920 to P.A.I.
PY - 2008/7/22
Y1 - 2008/7/22
N2 - Background: Studies show that high phosphotidylinositol 3,4,5-trisphosphate (PIP3) promotes cytoskeletal rearrangements and alters cell motility and chemotaxis, possibly through activation of protein kinase Bs (PKBs). However, chemotaxis can still occur in the absence of PIP3, and the identities of the PIP3-independent pathways remain unknown. Results: Here, we outline a PIP3-independent pathway linking temporal and spatial activation of PKBs by Tor complex 2 (TorC2) to the chemotactic response. Within seconds of stimulating Dictyostelium cells with chemoattractant, two PKB homologs, PKBA and PKBR1, mediate transient phosphorylation of at least eight proteins, including Talin, PI4P 5-kinase, two Ras GEFs, and a RhoGap. Surprisingly, all of the substrates are phosphorylated with normal kinetics in cells lacking PI 3-kinase activity. Cells deficient in TorC2 or PKB activity show reduced phosphorylation of the endogenous substrates and are impaired in chemotaxis. The PKBs are activated through phosphorylation of their hydrophobic motifs via TorC2 and subsequent phosphorylation of their activation loops. These chemoattractant-inducible events are restricted to the cell's leading edge even in the absence of PIP3. Activation of TorC2 depends on heterotrimeric G protein function and intermediate G proteins, including Ras GTPases. Conclusions: The data lead to a model where cytosolic TorC2, encountering locally activated small G protein(s) at the leading edge of the cell, becomes activated and phosphorylates PKBs. These in turn phosphorylate a series of signaling and cytoskeletal proteins, thereby regulating directed migration.
AB - Background: Studies show that high phosphotidylinositol 3,4,5-trisphosphate (PIP3) promotes cytoskeletal rearrangements and alters cell motility and chemotaxis, possibly through activation of protein kinase Bs (PKBs). However, chemotaxis can still occur in the absence of PIP3, and the identities of the PIP3-independent pathways remain unknown. Results: Here, we outline a PIP3-independent pathway linking temporal and spatial activation of PKBs by Tor complex 2 (TorC2) to the chemotactic response. Within seconds of stimulating Dictyostelium cells with chemoattractant, two PKB homologs, PKBA and PKBR1, mediate transient phosphorylation of at least eight proteins, including Talin, PI4P 5-kinase, two Ras GEFs, and a RhoGap. Surprisingly, all of the substrates are phosphorylated with normal kinetics in cells lacking PI 3-kinase activity. Cells deficient in TorC2 or PKB activity show reduced phosphorylation of the endogenous substrates and are impaired in chemotaxis. The PKBs are activated through phosphorylation of their hydrophobic motifs via TorC2 and subsequent phosphorylation of their activation loops. These chemoattractant-inducible events are restricted to the cell's leading edge even in the absence of PIP3. Activation of TorC2 depends on heterotrimeric G protein function and intermediate G proteins, including Ras GTPases. Conclusions: The data lead to a model where cytosolic TorC2, encountering locally activated small G protein(s) at the leading edge of the cell, becomes activated and phosphorylates PKBs. These in turn phosphorylate a series of signaling and cytoskeletal proteins, thereby regulating directed migration.
KW - CELLBIO
KW - SIGNALING
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U2 - 10.1016/j.cub.2008.06.068
DO - 10.1016/j.cub.2008.06.068
M3 - Article
C2 - 18635356
AN - SCOPUS:47049110588
SN - 0960-9822
VL - 18
SP - 1034
EP - 1043
JO - Current Biology
JF - Current Biology
IS - 14
ER -