I. Rubio et R. Wetzker, A permissive function of phosphoinositide 3-kinase in Ras activation mediated by inhibition of GTPase-activating proteins, CURR BIOL, 10(19), 2000, pp. 1225-1228
The activation status of the guanosine triphosphate (GTP)-binding protein P
as is dictated by the relative intensities of two opposing reactions: the f
ormation of active Ras-GTP complexes, promoted by guanine nucleotide exchan
ge factors (GEFs), and their conversion to inactive Ras-GDP as a result of
the deactivating action of GTPase-activating proteins (GAPs), The relevance
of phosphoinositide 3-kinase (PI 3-kinase) to these processes is still unc
lear. We have investigated the regulation of Pas activation by PI 3-kinase
in the myelomonocytic U937 cell line. These cells exhibited basal levels of
Ras-GTP, which were suppressed by two PI 3-kinase inhibitors and a dominan
t-negative PI 3-kinase, In addition, PI 3-kinase inhibition aborted Pas act
ivation by all stimuli tested, including foetal calf serum (FCS) and phorbo
l 12-myristate 13-acetate (TPA), Significantly, TPA does not activate PI 3-
kinase in U937 cells, indicating that PI 3 kinase has a permissive rather t
han an intermediary role in Pas activation, Investigation of the mechanism
of PI 3 kinase action revealed that inhibition of PI 3 kinase does not affe
ct nucleotide exchange on Pas but abrogates Ras-GTP accumulation through an
increase in GAP activity. These findings establish blockage of GAP action
as the mechanism underlying a permissive function of PI 3-kinase in Pas act
ivation. (C) 2000 Elsevier Science Ltd. All rights reserved.