Interleukin-3 and bryostatin 1 mediate rapid nuclear envelope protein phosphorylation in growth factor-dependent FDC-P1 hematopoietic cells. A possible role for nuclear protein kinase C.
Interleukin-3 (IL-3) is a lymphokine which stimulates the proliferation of normal and transformed multilineage hematopoietic cells. Recently we reported that bryostatin 1, a macrocyclic lactone and potent activator of protein kinase C, could stimulate normal multipotential hematopoietic progenitor cells in vitro in the absence of added polypeptide growth factors. We have now used the murine IL-3-dependent cell line FDC-P1, derived from normal murine marrow cells, to examine the early biochemical events associated with stimulation of hematopoietic cells. We find that both IL-3 and bryostatin 1 are mitogenic and stimulate the growth of FDC-P1 cells. Cells grown for extended periods in the presence of bryostatin 1 (1 nM) alone retain IL-3 responsiveness, indicating that bryostatin 1 does not induce an IL-3-independent state. Protein phosphorylation studies in cells treated with either IL-3 or bryostatin 1 indicate that both stimulators can mediate the rapid (within 5 min) serine-specific phosphorylation of several nuclear envelope polypeptides, including lamin B. Both IL-3- and bryostatin 1-mediated nuclear envelope phosphorylation is dose-dependent, occurring at concentrations which are mitogenic to FDC-P1 cells. The extent of nuclear envelope phosphorylation mediated by IL-3 and bryostatin 1 correlates with the mitogenic response. Furthermore, both mitogens mediate the rapid immunologic translocation of protein kinase C to the nuclear envelope where phosphorylation occurs. These data indicate that the early mitogenic signal(s) generated by IL-3 and bryostatin 1 may converge at the level of the nuclear envelope, perhaps through a protein kinase C-like activity which mediates phosphorylation of specific nuclear envelope polypeptides such as lamin B.
Fields
A P
AP
Johns Hopkins Oncology Center, Baltimore, Maryland 21231.
Pincus
S M
SM
Kraft
A S
AS
May
W S
WS
D000818:Animals; D054713:Bryostatins; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D006133:Growth Substances / Q000494:pharmacology*; D007377:Interleukin-3 / Q000494:pharmacology*; D007700:Kinetics; D007783:Lactones / Q000494:pharmacology*; D034921:Lamin Type B; D034882:Lamins; D018942:Macrolides; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008934:Mitogens / Q000494:pharmacology*; D008970:Molecular Weight; D009685:Nuclear Envelope / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000302:isolation & purification; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*
D054713:Bryostatins; D006133:Growth Substances; D007377:Interleukin-3; D007783:Lactones; D034921:Lamin Type B; D034882:Lamins; D018942:Macrolides; D008565:Membrane Proteins; D008934:Mitogens; D009687:Nuclear Proteins; D010750:Phosphoproteins; C046785:bryostatin 1; D011493:Protein Kinase C