Changes in the patterns of protein phosphorylation associated with granulocytic and monocytic-induced differentiation of HL-60 cells.
Using two-dimensional gel electrophoresis we have analyzed the pattern of phosphorylated proteins in HL-60 leukemia cells and changes associated with their differentiation into granulocyte and monocyte-like cells. In undifferentiated cells 18 spots with MW ranging from 110 to 17, kDa were individualized with high resolution and reproducibility. Myelocytic differentiation induced by dimethyl sulfoxide (DMSO) and retinoic acid (RA) resulted in a decrease of the overall phosphorylation, the disappearance of two proteins of 42 and 17 kDa, and the appearance of one new acidic protein of 46-48 kDa. These changes seem to be specifically related to this differentiation pathway since they are not found in two HL-60 subclones resistant to DMSO or RA-induced differentiation. Monocytic differentiation induced by 1,25-dihydroxyvitamin D3 [1,25 (OH)2D3] and the combination of RA + 1-B-D-arabinofuranosyl cytosine (Ara-C) was associated with the appearance of 2 proteins of 68 kDa and 2 proteins of 80 kDa located in the acidic region of the gel. The protein of 17 kDa, when disappeared completely in granulocytic-like cells was present in monocytic cells, this suggesting that its phosphorylation state may be involved in the control of the differentiation pathway of HL-60 cells. Data concerning the effect of phorbol myristate acetate (PMA) and histamine on the level of phosphorylation of various proteins in HL-60 cells have also been obtained and discussed. Our results show that the myelocytic and monocytic phenotypes are characterized by a specific pattern of phosphoproteins involving both phosphorylation and dephosphorylation reactions.
Faille
A
A
Poirier
O
O
Turmel
P
P
Chomienne
C
C
Charron
D J
DJ
Abita
J P
JP
D002117:Calcitriol / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D003561:Cytarabine / Q000494:pharmacology; D004121:Dimethyl Sulfoxide; D004591:Electrophoresis, Polyacrylamide Gel; D006098:Granulocytes / Q000166:cytology*; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism*; D008970:Molecular Weight; D009000:Monocytes / Q000166:cytology*; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology
D011506:Proteins; D003561:Cytarabine; D014212:Tretinoin; D006632:Histamine; D002117:Calcitriol; D013755:Tetradecanoylphorbol Acetate; D004121:Dimethyl Sulfoxide