Microfilament assembly is required for antigen-receptor-mediated activation of human B lymphocytes.
The mechanisms responsible for initiating the conversion of globular to filamentous actin (assembly) after stimulation of B lymphocytes and the role of these cytoskeletal changes in cell activation are incompletely understood. We investigated the molecular basis of the signals leading to actin polymerization and concentrated on the involvement of guanosine triphosphate (GTP)-binding regulatory proteins, and protein kinase C (PKC). In addition, we related these early events to later events in B-cell activation, including cell proliferation. Cross-linking the Ag receptor with Staphylococcus aureus Cowan I (SAC) or anti-IgM antibodies, or stimulation of PKC with phorbol ester induced a time- and concentration-dependent increase in the filamentous actin content of B cells. Inhibition or depletion of PKC resulted in decreased actin assembly induced by anti-IgM, SAC, and PMA, suggesting that the signal for polymerization is generated distally to PKC activation. Pertussis toxin pretreatment inhibited the responses to anti-IgM and SAC but not PMA, and direct stimulation of permeabilized cells with GTP gamma S induced microfilament assembly, indicating the involvement of a GTP-binding protein for receptor-mediated events. Disruption of actin polymerization with botulinum C2 toxin or cytochalasin D inhibited the assembly of actin and [3H]TdR incorporation induced by all stimuli. We conclude that human B cell activation by receptor-mediated stimuli results in actin polymerization by signaling pathways coupled to GTP-binding proteins. These changes in the cytoskeleton may be involved in the transduction of messages leading to responses such as proliferation in B lymphocytes.
Melamed
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Department of Pediatrics, Hospital for Sick Children, Toronto, Ontario, Canada.
Downey
G P
GP
Aktories
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Roifman
C M
CM
D008841:Actin Cytoskeleton / Q000502:physiology*; D000199:Actins / Q000378:metabolism*; D001402:B-Lymphocytes / Q000276:immunology*; D001905:Botulinum Toxins / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000502:physiology; D006801:Humans; D007075:Immunoglobulin M / Q000276:immunology; D008213:Lymphocyte Activation; D037342:Pertussis Toxin; D011493:Protein Kinase C / Q000502:physiology; D011947:Receptors, Antigen, B-Cell / Q000502:physiology; D013211:Staphylococcus aureus / Q000276:immunology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D000199:Actins; D007075:Immunoglobulin M; D011947:Receptors, Antigen, B-Cell; D010566:Virulence Factors, Bordetella; D015638:Cytochalasin D; D037342:Pertussis Toxin; D011493:Protein Kinase C; D001905:Botulinum Toxins; D019204:GTP-Binding Proteins; D013755:Tetradecanoylphorbol Acetate