Regulatory role of diacylglycerol kinase gamma in macrophage differentiation of leukemia cells.
Although nine diacylglycerol kinase (DGK) isozymes have been identified, our knowledge of their individual functions is still limited. Here we report that the levels of DGKgamma mRNA/protein in human leukemia HL-60 and U937 cells were rapidly and markedly decreased upon cellular differentiation into macrophages. In contrast, the enzyme expression remained almost unchanged in granulocytic differentiation pathway. Interestingly, the overexpression of wild-type or constitutively active DGKgamma, but not its kinase-dead mutant, markedly inhibited phorbol ester-induced cell attachment and nonspecific esterase activity, which are hallmarks of macrophage differentiation. We noted in this case that no effects were observed for the corresponding constructs of a closely related isozyme, DGKalpha. Prior to the cell attachment, phorbol ester induced translocation of DGKgamma from the cytoplasm to the cell periphery, resulting in its co-localization with F-actin together with protein kinase Cdelta. The results suggest that DGKgamma negatively regulates macrophage differentiation through its catalytic action operating on the cytoskeleton.
Department of Liberal Arts and Sciences, School of Health Sciences, Sapporo Medical University, South-1, West-17, Chuo-ku, Sapporo 060-8556, Japan. oyama@sapmed.ac.jp
Keiko
K
Yamada
Fumio
F
Sakane
Shin ichi
Si
Imai
Shuichi
S
Tsushima
Tomohiro
T
Murakami
Hideo
H
Kanoh
D000199:Actins / Q000032:analysis; D002448:Cell Adhesion; D002454:Cell Differentiation; D019852:Diacylglycerol Kinase / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D018922:HL-60 Cells; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics; D007700:Kinetics; D008264:Macrophages / Q000737:chemistry / Q000201:enzymology*; D009154:Mutation; D011493:Protein Kinase C / Q000032:analysis; D051745:Protein Kinase C-delta; D012333:RNA, Messenger / Q000096:biosynthesis; D020298:U937 Cells
D000199:Actins; D007527:Isoenzymes; D012333:RNA, Messenger; D019852:Diacylglycerol Kinase; C497258:PRKCD protein, human; D011493:Protein Kinase C; D051745:Protein Kinase C-delta