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Am J Physiol Renal Physiol 295: F1422-F1430, 2008. First published August 27, 2008; doi:10.1152/ajprenal.90310.2008 Free Article
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Localization of phosphatidylinositol phosphate kinase II{gamma} in kidney to a membrane trafficking compartment within specialized cells of the nephron

Jonathan H. Clarke,1 Piers C. Emson,2 and Robin F. Irvine1

1Department of Pharmacology, University of Cambridge and 2Laboratory of Molecular Neuroscience, Babraham Institute, Cambridge, United Kingdom

Submitted 15 May 2008 ; accepted in final form 24 August 2008

PIP4Ks (type II phosphatidylinositol 4-phosphate kinases) are phosphatidylinositol 5-phosphate (PtdIns5P) 4-kinases, believed primarily to regulate cellular PtdIns5P levels. In this study, we investigated the expression, localization, and associated biological activity of the least-studied PIP4K isoform, PIP4K{gamma}. Quantitative RT-PCR and in situ hybridization revealed that compared with PIP4K{alpha} and PIP4Kβ, PIP4K{gamma} is expressed at exceptionally high levels in the kidney, especially the cortex and outer medulla. A specific antibody was raised to PIP4K{gamma}, and immunohistochemistry with this and with antibodies to specific kidney cell markers showed a restricted expression, primarily distributed in epithelial cells in the thick ascending limb and in the intercalated cells of the collecting duct. In these cells, PIP4K{gamma} had a vesicular appearance, and transfection of kidney cell lines revealed a partial Golgi localization (primarily the matrix of the cis-Golgi) with an additional presence in an unidentified vesicular compartment. In contrast to PIP4K{alpha}, bacterially expressed recombinant PIP4K{gamma} was completely inactive but did have the ability to associate with active PIP4K{alpha} in vitro. Overall our data suggest that PIP4K{gamma} may have a function in the regulation of vesicular transport in specialized kidney epithelial cells.

phosphoinositide; phosphatidylinositol 5-phosphate 4-kinase; collecting duct; Golgi apparatus


Address for reprint requests and other correspondence: J. H. Clarke, Dept. of Pharmacology, Univ. of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK (e-mail: jhc30{at}cam.ac.uk)






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