Changes in phosphoproteins during commitment of HL60 cells to monocyte differentiation: evidence for multiple protein kinase involvement.
The human promyeloid cell line HL60 differentiates toward monocytes when treated with TPA. We have analyzed, by two-dimensional gel electrophoresis, the phosphoprotein patterns within HL60 cells, labeled to equilibrium with [32P]orthophosphate when cells were treated with suboptimal (1 nM), optimal (5 and 10 nM), and supraoptimal (40 and 100 nM) concentrations of 12-O-tetradecanylphorbol-13-acetate (TPA) as regards the induction of differentiation. No change was detected in the phosphoprotein pattern at 1 nM TPA, whereas four phosphoproteins showed increased levels of phosphorylation at 5 and 10 nM TPA. When cells were treated with 40 and 100 nM TPA, in total eight and ten proteins, respectively, were phosphorylated, including the above four proteins. Two proteins were dephosphorylated when cells were treated with 40 and 100 nM TPA. A 15-kd protein, phosphorylated when HL60 cells were treated with 5 nM TPA, was observed as an intense spot in autoradiographs of total cellular phosphoproteins of two variant HL60 cell lines that are unable to differentiate toward monocytes and prior to treatment with TPA. In the case of three variant cell lines, which like HL60 differentiate toward monocytes, the phosphoprotein spot was almost absent. Thus, paradoxically, the 15-kd phosphoprotein is affected by TPA although its constitutive level of expression or increased phosphorylation state is inversely related to the potential for monocyte differentiation. This observation, together with the TPA dose-response effects on protein phosphorylation, is discussed in relation to multiple protein kinase involvement.
Lord
J M
JM
Department of Immunology, Medical School, University of Birmingham, Edgbaston, UK.
Wong
A K
AK
Brown
G
G
D002454:Cell Differentiation / Q000187:drug effects; D006801:Humans; D007951:Leukemia, Myeloid / Q000201:enzymology / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000201:enzymology / Q000473:pathology*; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D010750:Phosphoproteins / Q000096:biosynthesis*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000201:enzymology / Q000473:pathology
D009363:Neoplasm Proteins; D010750:Phosphoproteins; D011494:Protein Kinases; D013755:Tetradecanoylphorbol Acetate