We assessed the roles of insulin receptor substrate-1 (IRS-I) and She in in
sulin action on farnesyltransferase (FTase) and geranylgeranyltransferase I
(GGTase I) using Chinese hamster ovary (CHO) cells that overexpress wild-ty
pe human insulin receptors (CHO-hIR-WT) or mutant insulin receptors lacking
the NPEY domain (CHO-Delta NPEY) or 3T3-L1 fibroblasts transfected with ad
enoviruses that express the PTB or SAIN domain of IRS-1 and She, the plecks
trin homology (PH) domain of IRS-1, or the Src homology 2 (SH2) domain of S
he. Insulin promoted phosphorylation of the alpha -subunit of FTase and GGT
ase I in CHO-hIR-WT cells, but was without effect in CHO-Delta NPEY cells.
Insulin increased FTase and GGTase I activities and the amounts of prenylat
ed Pas and RhoA proteins in CHO-hIR-WT (but not CHO-Delta NPEY) cells. Over
expression of the PTB or SAIN domain of IRS-l (which blocked both IRS-1 and
She signaling) prevented insulin-stimulated phosphorylation of the FTase a
nd GGTase I alpha -subunit activation of FTase and GGTase I and subsequent
increases in prenylated Res and RhoA proteins. In contrast, overexpression
of the IRS-1 PH domain, which impairs IRS-1 (but not She) signaling, did no
t alter insulin action on the prenyltransferases, but completely inhibited
the insulin effect on the phosphorylation of IRS-1 and on the activation of
phosphatidylinositol 3-kinase and Akt. Finally, overexpression of the Shc
SH2 domain completely blocked the insulin effect on FTase and GGTase I acti
vities without interfering with insulin signaling to MAPK. These data sugge
st that insulin signaling from its receptor to the prenyltransferases FTase
and GGTase I is mediated by the She pathway, but not the IRS-1/phosphatidy
linositol 3-kinase pathway. She-mediated insulin signaling to MAPK may be n
ecessary (but not sufficient) for activation of prenyltransferase activity,
An additional pathway involving the She SH2 domain may be necessary to med
iate the insulin effect on FTase and GGTase I.