Repor t Antigen - induced B Lymphocyte Activation Involves the p 21 ~ and ras . GAP Signaling Pathway

Ligation of a B lymphocyte surface immunoglobulin (slg) antigen receptor (AgR) by its specific Ag ligand initiates a signaling pathway that culminates in B cell activation. However, many events of this pathway have not been elucidated. Here we present three novel findings that demonstrate directly that AgR-mediated signaling in B cells functions by the p21ras/ras.GAP-dependent pathway. First, stimulation of TA3 7.9 Ag-specific murine B lymphoma cells for 2 rain with either Ag or F(ab')2 anti-IgM induces p21 ~ activation as measured by an increase in the GTP/GDP ratio of its bound nucleotides. This activation of p21 r" does not occur via a change in its guanine nucleotide exchange rate. Second, Ag stimulation results in the inhibition of activity of p120 ras.GAP, a protein that regulates p21 ~"~ activation. Tyrosine phosphorylation of ras.GAP occurs within 1 min after Ag stimulation but is no longer detectable at 20 rain after stimulation, at which time ras.GAP activity remains inhibited. Thus, tyrosine phosphorylation of ras.GAP is not required for the inhibition of its activity. Third, despite the role proposed for a ras.GAP-assodated p190 protein in the control of ras.GAP activity in B cells, p190 was not detectable either in antiras.GAP immunoprecipitates of [3SS]methionine labeled lysates of Ag-stimulated or -unstimulated 7.9 cells or as a tyrosine phosphoprotein in Western blots of anti-ras.GAP immunoprecipitates of Ag-stimulated 7.9 cell lysates. Inasmuch as the TA3 7.9 B lymphoma is representative of a mature, slgM-bearing B cell, our observations raise the intriguing possibility that the capacity of p190 to associate with ras.GAP and regulate the activities of ras.GAP and p21TM in a B cell is dependent on the stage of differentiation of the B cell.