caa a22 4500 467893845 CHVBK 20180406152756.0 cr unu---uuuuu 170328e20060301xx s 000 0 eng 10.1007/s11103-005-5548-x doi (NATIONALLICENCE)springer-10.1007/s11103-005-5548-x The Highly Charged Region of Plant β -type Phosphatidylinositol 4-kinase is Involved in Membrane Targeting and Phospholipid Binding [Elektronische Daten] [Ying Lou, Hui Ma, Wen-Hui Lin, Zhao-Qing Chu, Bernd Mueller-Roeber, Zhi-Hong Xu, Hong-Wei Xue] In Arabidopsis thaliana and Oryza sativa, two types of PI 4-kinase (PI4Ks) have been isolated and functionally characterized. The α-type PI4Ks (~220kDa) contain a PH domain, which is lacking in β-type PI4Ks (~120kDa). β-Type PI4Ks, exemplified by Arabidopsis AtPI4Kβ and rice OsPI4K2, contain a highly charged repetitive segment designated PPC (Plant PI4K Charged) region, which is an unique domain only found in plant β-type PI4Ks at present. The PPC region has a length of ~300 amino acids and harboring 11 (AtPI4Kβ) and 14 (OsPI4K2) repeats, respectively, of a 20-aa motif. Studies employing a modified yeast-based "Sequence of Membrane-Targeting Detection” system demonstrate that the PPC(OsPI4K2) region, as well as the former 8 and latter 6 repetitive motifs within the PPC region, are able to target fusion proteins to the plasma membrane. Further detection on the transiently expressed GFP fusion proteins in onion epidermal cells showed that the PPC(OsPI4K2) region alone, as well as the region containing repetitive motifs 1-8, was able to direct GFP to the plasma membrane, while the regions containing less repetitive motifs, i.e. 6, 4, 2 or single motif(s) led to predominantly intracellular localization. Agrobacterium-mediated transient expression of PPC-GFP fusion protein further confirms the membrane-targeting capacities of PPC region. In addition, the predominant plasma membrane localization of AtPI4Kβ was mediated by the PPC region. Recombinant PPC peptide, expressed in E. coli, strongly binds phosphatidic acid, PI and PI4P, but not phosphatidylcholine, PI5P, or PI(4,5)P2 in vitro, providing insights into potential mechanisms for regulating sub-cellular localization and lipid binding for the plant β-type PI4Ks. Springer, 2006 GFP nationallicence membrane targeting nationallicence PI4K nationallicence PPC ( P lant P I4Ks C harged) region nationallicence signal transduction nationallicence yeast nationallicence PI : phosphatidylinositol nationallicence PI4P : PI 4-monophosphate nationallicence PI5P : PI 5-monophosphate nationallicence PI(4,5)P2 : PI 4,5-bisphosphate nationallicence PI4K : PI 4-kinase nationallicence PA : phosphatidic acid nationallicence PAGE : polyacrylamide gel electrophoresis nationallicence Lou Ying National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut Ma Hui National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut Lin Wen-Hui National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut Chu Zhao-Qing National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut Mueller-Roeber Bernd Institute for Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Street 25, D-14476, Haus 20, Golm, Germany aut Xu Zhi-Hong National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut Xue Hong-Wei National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut Plant Molecular Biology Kluwer Academic Publishers 60/5(2006-03-01), 729-746 0167-4412 60:5<729 2006 60 11103 https://doi.org/10.1007/s11103-005-5548-x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11103-005-5548-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lou Ying National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut NATIONALLICENCE 700 1- Ma Hui National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut NATIONALLICENCE 700 1- Lin Wen-Hui National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut NATIONALLICENCE 700 1- Chu Zhao-Qing National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut NATIONALLICENCE 700 1- Mueller-Roeber Bernd Institute for Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Street 25, D-14476, Haus 20, Golm, Germany aut NATIONALLICENCE 700 1- Xu Zhi-Hong National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut NATIONALLICENCE 700 1- Xue Hong-Wei National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Science (SiBS), Chinese Academy of Sciences; and Partner Group of Max-Planck-Institute of Molecular Plant Physiology (MPI-MP) on "Plant Molecular Physiology and Signal Transduction”, 300 Fenglin Road, 200032, Shanghai, P.R. China aut NATIONALLICENCE 773 0- Plant Molecular Biology Kluwer Academic Publishers 60/5(2006-03-01), 729-746 0167-4412 60:5<729 2006 60 11103 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer