Identification of a phosphatidylinositol 3-hydroxy kinase in plant cells: association with the cytoskeleton

Radiolabelling experiments have revealed that plant cells contain the two 3-phosphorylated phosphoinositides: PtdIns3P and PtdIns(3,4)P2 [Brearley and Hanke (1992) Biochem. J. 283, 255-260]. However, nothing is known about the enzymes involved in the metabolism of these plant 3-phosphorylated phosphoinositides. In this study we demonstrate the presence of a PtdIns 3-hydroxy kinase(s) in plant cells. This activity was enriched in the cytoskeletal fraction whereas only low levels of phosphoinositide 3-hydroxy kinase could be detected in plasma membranes and microsomal preparations. This cytoskeletal phosphoinositide 3-hydroxy kinase was found to be wortmannin insensitive and thus resembles PtdIns-specific 3-hydroxy kinases of which vps34p is one example.