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First published online January 17, 2003; 10.1105/tpc.006676

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The Plant Cell, Vol. 15, 449-463, February 2003, Copyright © 2003,
American Society of Plant Biologists

Arabidopsis Inositol Polyphosphate 6-/3-Kinase Is a Nuclear Protein That Complements a Yeast Mutant Lacking a Functional ArgR-Mcm1 Transcription Complex

Hui-Jun Xiaa, Charles Brearleyb, Stephan Elge1,a, Boaz Kaplanc, Hillel Frommc,d and Bernd Mueller-Roeber1,2,a

a Max-Planck-Institute of Molecular Plant Physiology, D-14424 Potsdam, Germany
b School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, United Kingdom
c Department of Plant Sciences, The Weizmann Institute of Science, Rehovot 76100, Israel
d Centre for Plant Sciences, Leeds Institute for Biotechnology and Agriculture, School of Biology, University of Leeds, Leeds LS2 9JT, United Kingdom

2 To whom correspondence should be addressed. E-mail bmr{at}rz.uni-potsdam.de; fax 49-331-977-2512

Inositol 1,4,5-trisphosphate 3-kinase, and more generally inositol polyphosphate kinases (Ipk), play important roles in signal transduction in animal cells; however, their functions in plant cells remain to be elucidated. Here, we report the molecular cloning of a cDNA (AtIpk2{beta}) from a higher plant, Arabidopsis. Arabidopsis AtIpk2{beta} is a 33-kD protein that exhibits weak homology (~25% identical amino acids) with Ipk proteins from animals and yeast and lacks a calmodulin binding site, as revealed by sequence analysis and calmodulin binding assays. However, recombinant AtIpk2{beta} phosphorylates inositol 1,4,5-trisphosphate to inositol 1,4,5,6-tetrakisphosphate and also converts it to inositol 1,3,4,5,6-pentakisphosphate [Ins(1,3,4,5,6)P5]. AtIpk2{beta} also phosphorylates inositol 1,3,4,5-tetrakisphosphate to Ins(1,3,4,5,6)P5. Thus, the enzyme is a D3/D6 dual-specificity inositol phosphate kinase. AtIpk2{beta} complements a yeast ARG82/IPK2 mutant lacking a functional ArgR-Mcm1 transcription complex. This complex is involved in regulating Arg metabolism–related gene expression and requires inositol polyphosphate kinase activity to function. AtIpk2{beta} was found to be located predominantly in the nucleus of plant cells, as demonstrated by immunolocalization and fusion to green fluorescent protein. RNA gel blot analysis and promoter–{beta}-glucuronidase reporter gene studies demonstrated AtIpk2{beta} gene expression in various organs tested. These data suggest a role for AtIpk2{beta} as a transcriptional control mediator in plants.




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