Phosphoinositide 3′-kinases constitute a family of lipid kinases implicated in signal transduction through tyrosine kinase receptors and heterotrimeric G protein-linked receptors. Phosphoinositide 3′-kinases that bind to the platelet-derived growth factor receptor are composed of two subunits: the p85 subunit acts as an adapter and couples the catalytic p110 subunit to the activated receptor. There are different isoforms of p85 as well as of p110, the individual roles of which have been elusive. Using microinjection of inhibitory antibodies specific for either p110(alpha) or p110(beta) we have investigated the involvement of the two p110 isoforms in platelet-derived growth factor- and insulin-induced actin reorganization in porcine aortic endothelial cells. We have found that antibodies against p110(alpha), but not antibodies against p110(beta), inhibit platelet-derived growth factor-stimulated actin reorganization, whereas the reverse is true for inhibition of insulin-induced actin reorganization. These data indicate that the two phosphoinositide 3′-kinase isoforms have distinct roles in signal transduction pathways induced by platelet-derived growth factor and insulin.
REFERENCES
Antonetti
D. A.
, Algenstaedt
P.
, Kahn
C. R.
(1996
). Insulin receptor substrate 1 binds two novel splice variants of the regulatory subunit of phosphatidylinositol 3-kinase in muscle and brain.
Mol. Cell Biol
16
, 2195
–2203
Ayscough
K. R.
(1998
). In vivo functions of actin-binding proteins.
Curr. Opin. Cell Biol
10
, 102
–111
Cantrell
D.
(1998
). Lymphocyte signalling: a coordinating role for Vav?.
Curr. Biol
8
, 535
–538
Carpenter
C. L.
, Auger
K. R.
, Chanudhuri
M.
, Yoakim
M.
, Schaffhausen
B.
, Shoelson
S.
, Cantley
L. C.
(1993
). Phosphoinositide 3-kinase is activated by phosphopeptides that bind to the SH2 domains of the 85-kDa subunit.
J. Biol. Chem
268
, 9478
–9483
Cheatham
B.
, Vlahos
C. J.
, Cheatham
L.
, Wang
L.
, Blenis
J.
, Kahn
C. R.
(1994
). Phosphatidylinositol 3-kinase activation is required for insulin stimulation of pp70 S6 kinase, DNA synthesis, and glucose transporter translocation.
Mol. Cell Biol
14
, 4902
–4911
Chinkers
M.
, McKanna
J. A.
, Cohen
S.
(1979
). Rapid induction of morphological changes in human carcinoma cells A-431 by epidermal growth factors.
J. Cell Biol
83
, 260
–265
Claesson-Welsh
L.
(1994
). Platelet-derived growth factor receptor signals.
J. Biol. Chem
269
, 32023
–6
Cohen
G. B.
, Ren
R.
, Baltimore
D.
(1995
). Modular binding domains in signal transduction proteins.
Cell
80
, 237
–248
Deora
A. A.
, Win
T.
, Vanhaesebroeck
B.
, Lander
H. M.
(1998
). A Redox-triggered Ras-Effector Interaction. Recruitment of phosphatidylinositol 3-kinase to ras by redox stress.
J. Biol. Chem
273
, 29923
–29928
Fantl
W. J.
, Escobedo
J. A.
, Martin
G. A.
, Turck
C. W.
, del Rosario
M.
, McCormick
F.
, Williams
L. T.
(1992
). Distinct phosphotyrosines on a growth factor receptor bind to specific molecules that mediate different signaling pathways.
Cell
69
, 413
–423
Frevert
E. U.
, Kahn
B. B.
(1997
). Differential effects of constitutively active phosphatidylinositol 3-kinase on glucose transport, glycogen synthase activity, and DNA synthesis in 3T3-L1 adipocytes.
Mol. Cell Biol
17
, 190
–198
Fry
M. J.
(1994
). Structure, regulation and function of phosphoinositide 3-kinases.
Biochim. Biophys. Acta
1226
, 237
–268
Goldschmidt-Clermont
P. J.
, Machesky
L. M.
, Baldassare
J. J.
, Pollard
T. D.
(1990
). The actin-binding protein profilin binds to PIP2 and inhibits its hydrolysis by phospholipase C.
Science
247
, 1575
–1578
Goldschmidt-Clermont
P. J.
, Kim
J. W.
, Machesky
L. M.
, Rhee
S. G.
, Pollard
T. D.
(1991
). Regulation of phospholipase C-gamma 1 by profilin and tyrosine phosphorylation.
Science
251
, 1231
–1233
Guinebault
C.
, Payrastre
B.
, Racaud-Sultan
C.
, Mazarguil
H.
, Breton
M.
, Mauco
G.
, Plantavid
M.
, Chap
H.
(1995
). Integrin-dependent translocation of phosphoinositide 3-kinase to the cytoskeleton of thrombin-activated platelets involves specific interactions of p85with actin filaments and focal adhesion kinase.
J. Cell Biol
129
, 831
–842
Hartwig
J. H.
, Bokoch
G. M.
, Carpenter
C. L.
, Janmey
P. A.
, Taylor
L. A.
, Toker
A.
, Stossel
T. P.
(1995
). Thrombin receptor ligation and activated Rac uncap actin filament barbed ends through phosphoinositide synthesis in permeabilized human platelets.
Cell
82
, 643
–653
Heldin
C. H.
, Östman
A.
, Rönnstrand
L.
(1998
). Signal transduction via platelet-derived growth factor receptors.
Biochim. Biophys. Acta
1378
, 79
–113
Hiles
I. D.
, Otsu
M.
, Volinia
S.
, Fry
M. J.
, Gout
I.
, Dhand
R.
, Panayotou
G.
, Ruiz-Larrea
F.
, Thompson
A.
, Totty
N. F.
, Hsuan
J. J.
, Courtneidge
S. A.
, Parker
P. J.
, Waterfield
M. D.
(1992
). Phosphatidylinositol 3-kinase: structure and expression of the 110 kd catalytic subunit.
Cell
70
, 419
–429
Höglund
A. S.
, Karlsson
R.
, Arro
E.
, Fredriksson
B. A.
, Lindberg
U.
(1980
). Visualization of the peripheral weave of microfilaments in glia cells.
J. Muscle Res. Cell Motil
1
, 127
–146
Hooshmand-Rad
R.
, Claesson-Welsh
L.
, Wennström
S.
, Yokote
K.
, Siegbahn
A.
, Heldin
C. H.
(1997
). Involvement of phosphatidylinositide 3-kinase and Rac in platelet-derived growth factor-induced actin reorganization and chemotaxis.
Exp. Cell Res
234
, 434
–441
Hu
P.
, Mondino
A.
, Skolnik
E. Y.
, Schlessinger
J.
(1993
). Cloning of a novel, ubiquitously expressed human phosphatidylinositol 3-kinase and identification of its binding site on p85.
Mol. Cell Biol
13
, 7677
–7688
Hunter
S.
, Koch
B. L.
, Anderson
S. M.
(1997
). Phosphorylation of cbl after stimulation of Nb2 cells with prolactin and its association with phosphatidylinositol 3-kinase.
Mol. Endocrinol
11
, 1213
–1222
Inukai
K.
, Funaki
M.
, Ogihara
T.
, Katagiri
H.
, Kanda
A.
, Anai
M.
, Fukushima
Y.
, Hosaka
T.
, Suzuki
M.
, Shin
B. C.
, Takata
K.
, Yazaki
Y.
, Kikuchi
M.
, Oka
Y.
, Asano
T.
(1997
). p85gene generates three isoforms of regulatory subunit for phosphatidylinositol 3-kinase (PI 3-Kinase), p50, p55, and p85, with different PI3-kinase activity elevating responses to insulin.
J. Biol. Chem
272
, 7873
–7882
Jhun
B. H.
, Rose
D. W.
, Seely
B. L.
, Rameh
L.
, Cantley
L.
, Saltiel
A. R.
, Olefsky
J. M.
(1994
). Microinjection of the SH2 domain of the 85-kilodalton subunit of phosphatidylinositol 3-kinase inhibits insulin-induced DNA synthesis and c-fos expression.
Mol. Cell Biol
14
, 7466
–7475
Kaliman
P.
, Canicio
J.
, Shepherd
P. R.
, Beeton
C. A.
, Testar
X.
, Palacin
M.
, Zorzano
A.
(1998
). Insulin-like growth factors require phosphatidylinositol 3-kinase to signal myogenesis: dominant negative p85 expression blocks differentiation of L6E9 muscle cells.
Mol. Endocrinol
12
, 66
–77
Kazlauskas
A.
, Kashishian
A.
, Cooper
J. A.
, Valius
M.
(1992
). GTPase-activating protein and phosphatidylinositol 3-kinase bind to distinct regions of the platelet-derived growth factor receptor beta subunit.
Mol. Cell Biol
12
, 2534
–2544
Kerouz
N. J.
, Horsch
D.
, Pons
S.
, Kahn
C. R.
(1997
). Differential regulation of insulin receptor substrates-1 and-2 (IRS-1 and IRS-2) and phosphatidylinositol 3-kinase isoforms in liver and muscle of the obese diabetic (ob/ob) mouse.
J. Clin. Invest
100
, 3164
–3172
Klippel
A.
, Escobedo
J. A.
, Hirano
M.
, Williams
L. T.
(1994
). The interaction of small domains between the subunits of phosphatidylinositol 3-kinase determines enzyme activity.
Mol. Cell Biol
14
, 2675
–2685
Kotani
K.
, Yonezawa
K.
, Hara
K.
, Ueda
H.
, Kitamura
Y.
, Sakaue
H.
, Ando
A.
, Chavanieu
A.
, Calas
B.
, Grigorescu
F.
, Nishiyama
M.
, Waterfield
M. D.
, Kasuga
M.
(1994
). Involvement of phosphoinositide 3-kinase in insulin-or IGF-induced membrane ruffling.
EMBO J
13
, 2313
–2321
Kundra
V.
, Escobedo
J. A.
, Kazlauskas
A.
, Kim
H. K.
, Rhee
S. G.
, Williams
L. T.
, Zetter
B. R.
(1994
). Regulation of chemotaxis by the platelet-derived growth factor receptor-beta.
Nature
367
, 474
–476
Kurosu
H.
, Maehama
T.
, Okada
T.
, Yamamoto
T.
, Hoshino
S.
, Fukui
Y.
, Ui
M.
, Hazeki
O.
, Katada
T.
(1997
). Heterodimeric phosphoinositide 3-kinase consisting of p85 and p110is synergistically activated by the subunits of G proteins and phosphotyrosyl peptide.
J. Biol. Chem
272
, 24252
–24256
Lassing
I.
, Lindberg
U.
(1985
). Specific interaction between phosphatidylinositol 4, 5-bisphosphate and profilactin.
Nature
314
, 472
–474
Liu
X.
, Marengere
L. E.
, Koch
C. A.
, Pawson
T.
(1993
). The v-Src SH3 domain binds phosphatidylinositol 3-kinase.
Mol. Cell Biol
13
, 5225
–5232
Lu
P. J.
, Shieh
W. R.
, Rhee
S. G.
, Yin
H. L.
, Chen
C. S.
(1996
). Lipid products of phosphoinositide 3-kinase bind human profilin with high affinity.
Biochemistry
35
, 14027
–14034
McIlroy
J.
, Chen
D.
, Wjasow
C.
, Michaeli
T.
, Backer
J. M.
(1997
). Specific activation of p85-p110 phosphatidylinositol 3-kinase stimulates DNA synthesis by ras-and p70 S6 kinase-dependent pathways.
Mol. Cell Biol
17
, 248
–255
Mellström
K.
, Höglund
A. S.
, Nister
M.
, Heldin
C. H.
, Westermark
B.
, Lindberg
U.
(1983
). the effect of platelet-derived growth factor on morphology and motility of human glial cells.
J. Muscle Res. Cell Motil
4
, 589
–609
Nave
B. T.
, Haigh
R. J.
, Hayward
A. C.
, Siddle
K.
, Shepherd
P. R.
(1996
). Compartment-specific regulation of phosphoinositide 3-kinase by platelet-derived growth factor and insulin in 3T3-L1 adipocytes.
Biochem. J
318
, 55
–60
Nobes
C. D.
, Hawkins
P.
, Stephens
L.
, Hall
A.
(1995
). Activation of the small GTP-binding proteins rho and rac by growth factor receptors.
J. Cell Sci
108
, 225
–233
Pleiman
C. M.
, Hertz
W. M.
, Cambier
J. C.
(1994
). Activation of phosphatidylinositol-3kinase by Src-family kinase SH3 binding to the p85 subunit.
Science
263
, 1609
–1612
Roche
S.
, Koegl
M.
, Courtneidge
S. A.
(1994
). The phosphatidylinositol 3-kinaseis required for DNA synthesis induced by some, but not all, growth factors.
Proc. Nat. Acad. Sci. USA
91
, 9185
–9189
Roche
S.
, Downward
J.
, Raynal
P.
, Courtneidge
S. A.
(1998
). A function for phosphatidylinositol 3-kinase beta (p85-p110) in fibroblasts during mitogenesis: requirement for insulin-and lysophosphatidic acid-mediated signal transduction.
Mol. Cell Biol
18
, 7119
–7129
Shepherd
P. R.
, Nave
B. T.
, Rincon
J.
, Nolte
L. A.
, Bevan
A. P.
, Siddle
K.
, Zierath
J. R.
, Wallberg-Henriksson
H.
(1997
). Differential regulation of phosphoinositide 3-kinase adapter subunit variants by insulin in human skeletal muscle.
J. Biol. Chem
272
, 19000
–19007
Shibasaki
F.
, Fukami
K.
, Fukui
Y.
, Takenawa
T.
(1994
). Phosphatidylinositol 3-kinase binds to-actinin through the p85 subunit.
Biochem. J
302
, 551
–557
Siddhanta
U.
, McIlroy
J.
, Shah
A.
, Zhang
Y.
, Backer
J. M.
(1998
). Distinct roles for the p110and hVPS34 phosphatidylinositol 3 -kinases in vesicular trafficking, regulation of the actin cytoskeleton, and mitogenesis.
J. Cell Biol
143
, 1647
–1659
Singh
S. S.
, Chauhan
A.
, Murakami
N.
, Chauhan
V. P.
(1996
). Profilin and gelsolin stimulate phosphatidylinositol 3-kinase activity.
Biochemistry
35
, 16544
–16549
Small
J. V.
(1981
). Organization of actin in the leading edge of cultured cells: influence of osmium tetroxide and dehydration on the ultrastructure of actin meshworks.
J. Cell Biol
91
, 695
–705
Small
J. V.
, Rottner
K.
, Kaverina
I.
(1999
). Functional design in the actin cytoskeleton.
Curr. Opin. Cell Biol
11
, 54
–60
Stephens
L. R.
, Jackson
T. R.
, Hawkins
P. T.
(1993
). Agonist-stimulated synthesis of phosphatidylinositol(3,4,5)-trisphosphate: a new intracellular signalling system?.
Biochim. Biophys. Acta
1179
, 27
–75
Stephens
L.
, Smrcka
A.
, Cooke
F. T.
, Jackson
T. R.
, Sternweis
P. C.
, Hawkins
P. T.
(1994
). A novel phosphoinositide 3 kinase activity in myeloid-derived cells is activated by G proteinsubunits.
Cell
77
, 83
–93
Stoyanov
B.
, Volinia
S.
, Hanck
T.
, Rubio
I.
, Loubtchenkov
M.
, Malek
D.
, Stoyanova
S.
, Vanhaesebroeck
B.
, Dhand
R.
, Nurnberg
B.
, Gierschik
P.
, Seedorf
K.
, Hsuan
J. J.
, Waterfield
M. D.
, Wetzker
R.
(1995
). Cloning and characterization of a G protein-activated human phosphoinositide-3 kinase.
Science
269
, 690
–693
Tomiyama
K.
, Nakata
H.
, Sasa
H.
, Arimura
S.
, Nishio
E.
, Watanabe
Y.
(1995
). Wortmannin, a specific phosphatidylinositol 3-kinase inhibitor, inhibits adipocytic differentiation of 3T3-L1 cells.
Biochem. Biophys Res. Commun
212
, 263
–269
Vanhaesebroeck
B.
, Leevers
S. J.
, Panayotou
G.
, Waterfield
M. D.
(1997
). Phosphoinositide 3-kinases: a conserved family of signal transducers.
Trends Biochem. Sci
22
, 267
–272
Vanhaesebroeck
B.
, Welham
M. J.
, Kotani
K.
, Stein
R.
, Warne
P. H.
, Zvelebil
M. J.
, Higashi
K.
, Volinia
S.
, Downward
J.
, Waterfield
M. D.
(1997
). P110, a novel phosphoinositide 3-kinase in leukocytes.
Proc. Nat. Acad. Sci. USA
94
, 4330
–4335
Vanhaesebroeck
B.
, Higashi
K.
, Raven
C.
, Welham
M.
, Anderson
S.
, Brennan
P.
, Ward
S. G.
, Waterfield
M. D.
(1999
). Autophosphorylation of p110phosphoinositide 3-kinase: a new paradigm for the regulation of lipid kinases in vitro and in vivo.
EMBO J
18
, 1292
–1302
Wang
J.
, Auger
K. R.
, Jarvis
L.
, Shi
Y.
, Roberts
T. M.
(1995
). Direct association of Grb2 with the p85 subunit of phosphatidylinositol 3-kinase.
J. Biol. Chem
270
, 12774
–12780
Wang
Q.
, Bilan
P. J.
, Tsakiridis
T.
, Hinek
A.
, Klip
A.
(1998
). Actin filaments participate in the relocalization of phosphatidylinositol 3-kinase to glucose transporter-containing compartments and in the stimulation of glucose uptake in 3T3-L1 adipocytes.
Biochem. J
331
, 917
–928
Wennström
S.
, Hawkins
P.
, Cooke
F.
, Hara
K.
, Yonezawa
K.
, Kasuga
M.
, Jackson
T.
, Claesson-Welsh
L.
, Stephens
L.
(1994
). Activation of phosphoinositide 3-kinase is required for PDGF-stimulated membrane ruffling.
Curr. Biol
4
, 385
–393
Wennström
S.
, Siegbahn
A.
, Yokote
K.
, Arvidsson
A. K.
, Heldin
C. H.
, Mori
S.
, Claesson-Welsh
L.
(1994
). Membrane ruffling and chemotaxis transduced by the PDGF-receptor require the binding site for phosphatidylinositol 3 kinase.
Oncogene
9
, 651
–660
Yao
R.
, Cooper
G. M.
(1995
). Requirement for phosphatidylinositol-3 kinase in the prevention of apoptosis by nerve growth factor.
Science
267
, 2003
–2006
Yuan
Z. M.
, Utsugisawa
T.
, Huang
Y.
, Ishiko
T.
, Nakada
S.
, Kharbanda
S.
, Weichselbaum
R.
, Kufe
D.
(1997
). Inhibition of phosphatidylinositol 3-kinase by c-Abl in the genotoxic stress response.
J. Biol. Chem
272
, 23485
–23488
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