Translocation of protein phosphatase 1 catalytic subunits during 1,25-dihydroxyvitamin D3-induced monocytic differentiation of HL-60 cells.
To elucidate the roles of protein phosphatases type 1 (PP1) and type 2A (PP2A) in 1,25-dihydroxy-cholecalciferol [1,25(OH)2D3]-induced differentiation of HL-60 cells into monocytes, we examined the enzyme activity and the protein and gene expressions of PP1 and PP2A in these cells. Calyculin-A augmented the 1,25(OH)2D3-induced differentiation of the cells. Treatment of the cells with 1,25(OH)2D3 led to a decrease in PP1-like activity in the cytosol fraction, with a concomitant increase in the membrane and nuclear PP1-like activity, as determined when protein phosphatase activity was assayed using myosin light chain as substrate in the presence of 5 nM okadaic acid. Western blot analysis with antibodies specific for PP1 catalytic subunit isozymes (PP1 alpha, PP1 gamma, and PP1 delta) showed that all three PP1 isozymes were expressed but were differentially distributed in each cellular fraction. Subcellular redistribution of PP1-like activity during 1,25(OH)2D3-induced differentiation was mainly attributed to PP1 gamma and PP1 alpha proteins. In contrast, the localizations of PP1 delta and PP2A catalytic and regulatory subunits were not significantly affected by 1,25(OH)2D3 treatment. The gene expressions of PP1 alpha and PP1 gamma appeared to be constant during processes of monocytic differentiation. The correlation between phenotypic and functional changes of HL-60 cells on the one hand and subcellular redistribution of PP1-like activity on the other suggest that the translocations of PP1 alpha and PP1 gamma isozymes may contribute to the 1,25(OH)2D3-induced monocytic differentiation of HL-60 cells.
2nd Department of Internal Medicine, Mie University School of Medicine, Japan.
S B
SB
Omay
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H
Ogasawara
H
H
Toyoda
K
K
Nakai
H
H
Shima
M
M
Nagao
M C
MC
Mumby
H
H
Shiku
M
M
Nishikawa
D002117:Calcitriol / Q000494:pharmacology*; D002384:Catalysis; D002454:Cell Differentiation / Q000187:drug effects; D006801:Humans; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D015473:Leukemia, Promyelocytic, Acute / Q000188:drug therapy / Q000201:enzymology* / Q000473:pathology; D046911:Macromolecular Substances; D008387:Marine Toxins; D009000:Monocytes / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology*; D010080:Oxazoles / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D054645:Protein Phosphatase 1; D012333:RNA, Messenger / Q000235:genetics; D014178:Translocation, Genetic
D007527:Isoenzymes; D046911:Macromolecular Substances; D008387:Marine Toxins; D010080:Oxazoles; D012333:RNA, Messenger; C059041:calyculin A; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1; D002117:Calcitriol