Inositol phosphates are widely produced throughout animal and plant tissues. Diphosphoinositol pentakisphosphate (InsP7) contains an energetic pyrophosphate bond. Here we demonstrate that disruption of inositol hexakisphosphate kinase 1 (InsP6K1), one of the three mammalian inositol hexakisphosphate kinases (InsP6Ks) that convert inositol hexakisphosphate (InsP6) to InsP7, conferred enhanced phosphatidylinositol-(3,4,5)-trisphosphate (PtdIns(3,4,5)P3)-mediated membrane translocation of the pleckstrin homology domain of the kinase Akt and thus augmented downstream PtdIns(3,4,5)P3 signaling in mouse neutrophils. Consequently, these neutrophils had greater phagocytic and bactericidal ability and amplified NADPH oxidase–mediated production of superoxide. These phenotypes were replicated in human primary neutrophils with pharmacologically inhibited InsP6Ks. In contrast, an increase in intracellular InsP7 blocked chemoattractant-elicited translocation of the pleckstrin homology domain to the membrane and substantially suppressed PtdIns(3,4,5)P3-mediated cellular events in neutrophils. Our findings establish a role for InsP7 in signal transduction and provide a mechanism for modulating PtdIns(3,4,5)P3 signaling in neutrophils.
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We thank L. Silberstein, J. Manis and L. Chai for discussions. Supported by the US National Institutes of Health (HL085100, AI076471, HL092020 and GM076084 to H.R.L.).
The authors declare no competing financial interests.
Supplementary Figures 1–10 and Supplementary Methods (PDF 1117 kb)
Chemoattractant-elicited polarization of WT neutrophils (MOV 2006 kb)
Chemoattractant-elicited polarization of InsP6K−/− neutrophils (MOV 1696 kb)
Adhesion of WT (left) and InsP6K−/− (right) neutrophils under shear flow (MOV 799 kb)
Detachment of adhered WT (left) and InsP6K−/− (right) neutrophils under shear flow (MOV 1119 kb)
Chemotaxis of WT neutrophils (bottom) towards 1 AM of fMLP (top) in EZ-TAXIScan chamber (MOV 882 kb)
Chemotaxis of InsP6K1−/− neutrophils (bottom) towards 1 AM of fMLP (top) in EZ-TAXIScan chamber (MOV 862 kb)
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Prasad, A., Jia, Y., Chakraborty, A. et al. Inositol hexakisphosphate kinase 1 regulates neutrophil function in innate immunity by inhibiting phosphatidylinositol-(3,4,5)-trisphosphate signaling. Nat Immunol 12, 752–760 (2011). https://doi.org/10.1038/ni.2052
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