Carbohydrate-dependent signaling from the phosphatidylglucoside-based microdomain induces granulocytic differentiation of HL60 cells.
Glycosphingolipids form glycosphingolipid signaling microdomains. Here, we report an unrecognized type of phosphatidylglucoside (PhGlc)-based lipid microdomain in HL60 cells. Treatment of cells with rGL-7, which preferentially reacts with PhGlc, induced differentiation of HL60 cells. This was manifested by the appearance of nitroblue tetrazolium-positive cells together with CD38 expression and c-Myc down-regulation. We determined the molecular mechanisms underlying early stages of signal transduction. rGL-7 treatment induced rapid tyrosine phosphorylation of Src family protein kinases Lyn and Hck. Reduction of endogenous cholesterol after application of methyl-beta-cyclodextrin suppressed rGL-7-stimulated tyrosine phosphorylation. Phosphorylated proteins and PhGlc colocalized in the Triton X-100 insoluble, light buoyant density fraction after sucrose gradient ultracentrifugation of HL60 cell lysates. This suggests PhGlc-based microdomain is involved in GL-7 signaling. Ligation of known components of microdomains, such as sphingomyelin and ganglioside GM1, with corresponding antibodies failed to induce differentiation and tyrosine phosphorylation. These results show that PhGlc constitutes a previously undescribed lipid signaling domain, and the glucose residue of PhGlc is critical for organization of the carbohydrate-dependent signaling domain involved in cellular differentiation of HL60 cells.
Nagatsuka
Yasuko
Y
Department of Microbiology, Nihon University School of Medicine, Itabashi-ku, Tokyo 173-8610, Japan. ynaga@med.nihon-u.ac.jp
Hara-Yokoyama
Miki
M
Kasama
Takeshi
T
Takekoshi
Masataka
M
Maeda
Fumiko
F
Ihara
Seiji
S
Fujiwara
Shigeyoshi
S
Ohshima
Eriko
E
Ishii
Kumiko
K
Kobayashi
Toshihide
T
Shimizu
Kazufumi
K
Hirabayashi
Yoshio
Y
D036541:ADP-ribosyl Cyclase / Q000096:biosynthesis; D051997:ADP-ribosyl Cyclase 1; D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000096:biosynthesis; D050260:Carbohydrate Metabolism; D002454:Cell Differentiation; D003505:Cyclodextrins / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D015536:Down-Regulation; D005677:G(M1) Ganglioside / Q000378:metabolism; D020404:Glycerophospholipids / Q000737:chemistry / Q000378:metabolism*; D006098:Granulocytes / Q000166:cytology; D018922:HL-60 Cells; D006801:Humans; D015151:Immunoblotting; D050356:Lipid Metabolism; D008562:Membrane Glycoproteins; D021962:Membrane Microdomains / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D017830:Octoxynol / Q000494:pharmacology; D010641:Phenotype; D010766:Phosphorylation; D011233:Precipitin Tests; D015398:Signal Transduction; D013109:Sphingomyelins / Q000378:metabolism; D013997:Time Factors; D014212:Tretinoin / Q000378:metabolism / Q000494:pharmacology; D047392:beta-Cyclodextrins
D015703:Antigens, CD; D003505:Cyclodextrins; D003902:Detergents; D020404:Glycerophospholipids; D008562:Membrane Glycoproteins; D013109:Sphingomyelins; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; C429823:phosphatidylglucose; D005677:G(M1) Ganglioside; D014212:Tretinoin; D017830:Octoxynol; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1