M. Cormont et al., RAB4 IS PHOSPHORYLATED BY THE INSULIN-ACTIVATED EXTRACELLULAR-SIGNAL-REGULATED KINASE ERK1, European journal of biochemistry, 219(3), 1994, pp. 1081-1085
Rab4, a low-molecular-mass GTP-binding protein, is associated with ves
icles containing Glut 4 in adipocytes. Following insulin stimulation,
the translocation of Glut 4 to the plasma membrane is associated with
the movement of Rab4 to the cytosol. The same modifications are induce
d by the phosphatase inhibitor, okadaic acid. To establish a possible
role for phosphorylation in Rab4 cycling, we searched for insulin-stim
ulated cytosolic kinase(s) which could phosphorylate Rab4. In 3T3-L1 a
dipocytes, insulin induced a rapid and transient activation of cytosol
ic kinase(s), which phosphorylated Rab4 in vitro. At least part of the
Rab4 phosphorylation can be accounted for by ERK (extracellular-signa
l-regulated kinases) since immunopurified ERK1 from insulin-stimulated
cells phosphorylated Rab4 with a comparable time-course. Both with cy
tosolic extracts and immunopurified ERK1, only serine residues were ph
osphorylated on Rab4. The phosphorylation site was localized in the C-
terminus of the molecule, and occurred very probably on Ser196. These
results indicate that Rab4 is an in vitro substrate for ERK, and sugge
st that the insulin-induced movement of Rab4 from the Glut-4-containin
g vesicles to the cytosol could result from phosphorylation of Rab4 by
ERK.