Interleukin-3 and bryostatin-1 mediate hyperphosphorylation of BCL2 alpha in association with suppression of apoptosis.
Using murine myeloid factor-dependent FDC-P1/ER cells, we demonstrate that the hematopoietic growth factors interleukin-3 and erythropoietin and bryostatin-1, a macrocyclic lactone natural product and potent activator of protein kinase C (PKC), suppress apoptosis and induce the rapid serine phosphorylation of Bc12 alpha. Expression of recombinant wild type Bc12 alpha in NFS/N1.H-7 cells confirms that murine Bc12 alpha is phosphorylated following PKC activation. The PKC inhibitors H-7 and staurosporine, but not the protein kinase A inhibitor HA1004, block not only interleukin-3- and bryostatin-1-induced hyperphosphorylation of Bc12 alpha but also their anti-apoptotic effect on growth factor-dependent cells, suggesting a role for activated PKC in both processes. A potential direct role for a classic isoform of PKC is indicated by the Ca(2+)-dependent nature of phosphorylation of Bc12 alpha mediated by purified PKC in vitro. Comparative phosphopeptide maps confirm that Bc12 alpha phosphorylation occurs on identical serine site(s) whether phosphorylation occurs in cells following agonist treatment or directly by PKC in vitro. These findings strongly support a role for activated PKC in growth factor-induced Bc12 alpha phosphorylation as well as suppression of apoptosis.
May
W S
WS
Johns Hopkins Oncology Center, Johns Hopkins University School of Medicine, Baltimore, Maryland 21231.
Tyler
P G
PG
Ito
T
T
Armstrong
D K
DK
Qatsha
K A
KA
Davidson
N E
NE
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000470:Alkaloids / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000502:physiology*; D001483:Base Sequence; D054713:Bryostatins; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004921:Erythropoietin / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000502:physiology*; D007377:Interleukin-3 / Q000494:pharmacology*; D007546:Isoquinolines / Q000494:pharmacology; D007783:Lactones / Q000494:pharmacology*; D018942:Macrolides; D051379:Mice; D008934:Mitogens / Q000494:pharmacology; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010766:Phosphorylation; D010879:Piperazines / Q000494:pharmacology; D011233:Precipitin Tests; D011493:Protein Kinase C / Q000378:metabolism; D047428:Protein Kinase Inhibitors; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D019311:Staurosporine
D000470:Alkaloids; D000970:Antineoplastic Agents; D054713:Bryostatins; D007377:Interleukin-3; D007546:Isoquinolines; D007783:Lactones; D018942:Macrolides; D008934:Mitogens; D010879:Piperazines; D047428:Protein Kinase Inhibitors; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D011994:Recombinant Proteins; D004921:Erythropoietin; C046785:bryostatin 1; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011493:Protein Kinase C; D019311:Staurosporine