Abstract
Phosphatidylinositol-3 kinase (PI3K) activity is essential for normal B-cell receptor (BCR)-mediated responses. To understand the mechanisms of PI3K regulation during B-cell activation, we performed a series of biochemical analysis on primary B cells, and found that activity of Src family tyrosine kinases (SFK) is crucial for the activation of PI3K following BCR ligation and this is regulated by the SFK Lyn. We show that the hyperresponsive phenotype of B cells lacking Lyn is predicated on significantly increased basal and inducible PI3K activity that correlates with the constitutive hypophosphorylation of PAG/Cbp (phosphoprotein associated with glycosphingolipid-enriched microdomains/Csk-binding protein), a concomitant reduction in bound Csk in Lyn(-/-) B cells and elevated levels of active Fyn. Regulating SFK activity may thus be a central mechanism by which Lyn regulates PI3K activity in B cells. This study defines the molecular connection between the BCR and PI3K and reveals this to be a point of Lyn-mediated regulation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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B-Lymphocytes / immunology*
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B-Lymphocytes / metabolism
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Blotting, Western
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Enzyme Activation / immunology
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Female
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Male
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Membrane Proteins / immunology
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Membrane Proteins / metabolism
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Mitogen-Activated Protein Kinase 1 / immunology
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Mitogen-Activated Protein Kinase 1 / metabolism
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Models, Immunological
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NF-kappa B / immunology
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NF-kappa B / metabolism
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Phosphatidylinositol 3-Kinases / immunology*
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Phosphatidylinositol 3-Kinases / metabolism
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Phosphoproteins / immunology
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Phosphoproteins / metabolism
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Phosphorylation / immunology
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Proto-Oncogene Proteins c-akt / immunology
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Proto-Oncogene Proteins c-akt / metabolism
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Receptors, Antigen, B-Cell / immunology*
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Receptors, Antigen, B-Cell / metabolism
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src-Family Kinases / genetics
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src-Family Kinases / immunology*
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src-Family Kinases / metabolism
Substances
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Membrane Proteins
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NF-kappa B
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Pag1 protein, mouse
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Phosphoproteins
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Receptors, Antigen, B-Cell
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Phosphatidylinositol 3-Kinases
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lyn protein-tyrosine kinase
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src-Family Kinases
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Proto-Oncogene Proteins c-akt
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Mitogen-Activated Protein Kinase 1