Evidence that 3'-phosphorylated polyphosphoinositides are generated at the nuclear surface: use of immunostaining technique with monoclonal antibodies specific for PI 3,4-P(2).
Phosphatidylinositol (PI) 3,4-P(2) is a phosphoinositide that has been shown to be important for signal transduction in growth factor stimulation. We have produced monoclonal antibodies specific for PI 3,4-P(2), which were able to detect PI 3,4-P(2) generated in 293T cells treated with H(2)O(2), or in MKN45/BD110 cells expressing activated PI 3-kinase in immunostaining. Prolonged treatment with 0.05% Tween 20 resulted in detection of staining not only at the plasma membrane, but also at the nuclear surface, indicating that 3'-phosphorylated phosphoinositides can be generated and function in the nucleus.
Yokogawa
T
T
Department of Applied Biological Chemistry, Graduate School of Agriculture and Life Science, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, Japan.
Nagata
S
S
Nishio
Y
Y
Tsutsumi
T
T
Ihara
S
S
Shirai
R
R
Morita
K
K
Umeda
M
M
Shirai
Y
Y
Saitoh
N
N
Fukui
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007150:Immunohistochemistry; D008081:Liposomes / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009685:Nuclear Envelope / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000187:drug effects / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D008081:Liposomes; D018129:Phosphatidylinositol Phosphates; C060980:phosphatidylinositol 3,4-diphosphate; D006861:Hydrogen Peroxide; D019869:Phosphatidylinositol 3-Kinases