Vav is necessary for prolactin-stimulated proliferation and is translocated into the nucleus of a T-cell line.
Stimulation of the prolactin receptor (PRLr) with ligand activates multiple kinase cascades. The proximal mediators involved in the activation of the PRL-activated Raf-1 cascade in T-cells, however, remain poorly characterized. The role of one proximal signaling protein, namely p95vav, during PRLr signal transduction was examined in the Nb2 T-cell line. The novel results obtained here indicate that Vav is transiently associated with the PRLr and is necessary for PRL-stimulated proliferation. During PRL stimulation, a rapid and dramatic increase in guanine nucleotide exchange factor (GEF) activity was found to be associated with Vav immunoprecipitates. Concomitantly, an increase in Vav phosphorylation on serine-threonine residues was observed. The Vav-associated GEF activation could be inhibited by staurosporine and calphostin, but not herbimycin, suggesting a modulatory role for phosphorylation at serine-threonine residues. Treatment of Nb2 cells with antisense Vav oligonucleotide ablated Vav expression and blocked PRL-driven proliferation, but failed to inhibit PRL-induced GEF activation within Nb2 lysates. These data indicate that GEF activity may not be intrinsic to Vav as has been previously suggested, but either resides in or is complemented by an associated GEF. Subsequent to the transient activation of associated GEF activity, Vav was found to translocate into the Nb2 cell nucleus. Thus, Vav may utilize two independent mechanisms in T-cells, namely the activation of an associated GEF and direct nuclear internalization, to mediate PRLr signaling.
Clevenger
C V
CV
Department of Pathology, University of Pennsylvania Medical Center, Philadelphia 19104, USA.
Ngo
W
W
Sokol
D L
DL
Luger
S M
SM
Gewirtz
A M
AM
D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D018797:Cell Cycle Proteins; D002455:Cell Division / Q000187:drug effects / Q000502:physiology*; D002467:Cell Nucleus / Q000378:metabolism*; D018076:DNA, Complementary; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011388:Prolactin / Q000494:pharmacology*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D051567:Proto-Oncogene Proteins c-vav; D051381:Rats; D011981:Receptors, Prolactin / Q000378:metabolism*; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D018797:Cell Cycle Proteins; D018076:DNA, Complementary; D011518:Proto-Oncogene Proteins; D051567:Proto-Oncogene Proteins c-vav; D011981:Receptors, Prolactin; C496620:Vav1 protein, rat; D011388:Prolactin