Bistratene A: a novel compound causing changes in protein phosphorylation patterns in human leukemia cells.
Bistratene A, a polyether toxin isolated from the colonial ascidian Lissoclinum bistratum, causes incomplete differentiation of human leukemia (HL-60) cells apparently through a mechanism not involving protein kinase C. In view of the importance of phosphorylation/dephosphorylation in cellular growth and differentiation we have investigated protein phosphorylation in these cells following exposure to bistratene A, using two-dimensional polyacrylamide gel electrophoresis. Marked increases in the phosphorylation of a protein of 20 kDa, pl 6.7, and a basic protein of 25 kDa were observed after incubation with bistratene A. A comparison was made with cells treated with 12-O-tetradecanoylphorbol 13-acetate and bryostatin 5. While changes in phosphorylation patterns were observed with these two compounds, the 20 kDa and 25 kDa proteins did not undergo phosphorylation changes. The 20 kDa protein was induced rapidly by very low concentrations of bistratene A reaching near maximal levels with 10 nM at 15 min exposure. This protein was found to be localised to the cytoplasm. Phosphoaminoacid analysis demonstrated that the majority of 32P was present in serine and tyrosine residues. The increased phosphorylation of the 20 kDa protein appeared to be due to hyperphosphorylation of existing protein although there was some increase in the amount of the protein. These results suggest that bistratene A will be a useful tool with which to investigate cellular differentiation mechanisms.
Watters
D J
DJ
Queensland Cancer Fund Research Unit, Queensland Institute of Medical Research, Brisbane, Australia.
Michael
J
J
Hemphill
J E
JE
Hamilton
S E
SE
Lavin
M F
MF
Pettit
G R
GR
D000081:Acetamides; D000970:Antineoplastic Agents / Q000494:pharmacology; D054713:Bryostatins; D004988:Ethers, Cyclic / Q000494:pharmacology*; D006801:Humans; D007783:Lactones / Q000494:pharmacology; D007938:Leukemia; D018942:Macrolides; D009363:Neoplasm Proteins / Q000032:analysis / Q000187:drug effects* / Q000502:physiology; D010766:Phosphorylation / Q000187:drug effects; D011714:Pyrans; D013141:Spiro Compounds; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000081:Acetamides; D000970:Antineoplastic Agents; D054713:Bryostatins; D004988:Ethers, Cyclic; D007783:Lactones; D018942:Macrolides; D009363:Neoplasm Proteins; D011714:Pyrans; D013141:Spiro Compounds; C059975:bistratene A; C078062:bryostatin 5; D013755:Tetradecanoylphorbol Acetate