Differential requirement for Rho family GTPases in an oncogenic insulin-like growth factor-I receptor-induced cell transformation.
Insulin-like growth factor I receptor (IGFR) plays an important role in cell growth and transformation. We dissected the downstream signaling pathways of an oncogenic variant of IGFR, Gag-IGFR, called NM1. Loss of function mutants of NM1, Phe-1136 and dS2, that retain kinase activity but are attenuated in their transforming ability were used to identify signaling pathways that are important for transformation of NIH 3T3 cells. MAPK, phospholipase C gamma, and Stat3 were activated to the same extent by NM1 and its two mutants, suggesting that activation of these pathways, individually or in combination, was not sufficient for NM1-induced cell transformation. The mutant dS2 has decreased IRS-1 phosphorylation levels and IRS-1-associated phosphatidylinositol 3'-kinase activity, suggesting that this impairment may be in part responsible for the defectiveness of dS2. We show that Rho family members, RhoA, Rac1, and Cdc42 are activated by NM1, and this activation, particularly RhoA and Cdc42, is attenuated in both mutants of NM1. Dominant negative mutants of Rho, Rac, and Cdc42 inhibited NM1-induced cell transformation, as measured by focus and colony forming ability. Dominant negative Rho most potently inhibited the focus forming activity, whereas Cdc42 was most effective in inhibiting the colony forming ability of NM1-expressing cells. Conversely, constitutively activated (ca) Rho is more effective than ca Rac or ca Cdc42 in rescuing the focus forming ability of the mutants. By contrast, ca Cdc42 is most effective in rescuing the colony forming ability of both mutants.
Sachdev
P
P
Department of Microbiology, Mount Sinai School of Medicine, New York, New York 10029, USA.
Jiang
Y X
YX
Li
W
W
Miki
T
T
Maruta
H
H
Nur-E-Kamal
M S
MS
Wang
L H
LH
D016475:3T3 Cells; D000818:Animals; D002471:Cell Transformation, Neoplastic; D015972:Gene Expression Regulation, Neoplastic; D051379:Mice; D009154:Mutation; D017526:Receptor, IGF Type 1 / Q000502:physiology*; D015398:Signal Transduction; D020741:rho GTP-Binding Proteins / Q000502:physiology*
D017526:Receptor, IGF Type 1; D020741:rho GTP-Binding Proteins