PKC activity and protein phosphorylation in regulation of sIg mediated B cell activation.
The inhibitory and stimulatory elements of cellular signalling associated with activation of protein kinase C (PKC) in murine B lymphocytes were investigated by employing two PKC activators with opposing effects on cell proliferation. Being an inhibitor of anti-Ig mediated proliferation, the phorbol ester PDBU induced a more substantial translocation of cytosolic PKC activity than the alkaloid PKC activator indolactam, which enhances anti-Ig mediated B cell proliferation. PDBU and indolactam were equally effective kinase activators, as determined by 32P incorporation of the substrate proteins. Concentrations of indolactam which induced an inhibition of anti-Ig mediated B cell proliferation also induced a precipitous decline in detergent soluble cellular PKC activity, which was comparable with 1 microM PDBU. The induced phosphoprotein patterns were similar, with an exception of the nuclear envelope protein lamin B, which was prominently phosphorylated by PDBU but not by stimulatory concentrations of indolactam. The enhanced phosphorylation of lamin B was associated with cellular growth arrest: inhibitory concentrations of indolactam induced the phosphorylation of lamin B equal to PDBU, whereas an increased phosphorylation of lamin B was never observed upon stimulation with anti-Ig. Together, inhibition of anti-Ig mediated B cell proliferation was related to down-regulation of cytoplasmic PKC and induction of nuclear PKC-dependent phosphorylation.
Lindsberg
M L
ML
Department of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD 20814-4799.
Feuerstein
N
N
Balapure
A K
AK
Mond
J J
JJ
D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D007211:Indoles / Q000494:pharmacology*; D007769:Lactams / Q000494:pharmacology*; D008213:Lymphocyte Activation / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008811:Mice, Inbred DBA; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology*; D010750:Phosphoproteins / Q000378:metabolism*; D011493:Protein Kinase C / Q000187:drug effects / Q000378:metabolism*
D007211:Indoles; D007769:Lactams; D010750:Phosphoproteins; D015240:Phorbol 12,13-Dibutyrate; C043969:indolactam V; D011493:Protein Kinase C