First published online June 12, 2003; 10.1104/pp.103.019943
Plant Physiology 132:1623-1630 (2003)
© 2003 American Society of Plant Biologists
DEVELOPMENT AND HORMONE ACTION
PINOID-Mediated Signaling Involves Calcium-Binding Proteins
René Benjamins1,
Carlos S. Galván Ampudia,
Paul J.J. Hooykaas and
Remko Offringa*
Institute of Biology, Leiden University, Wassenaarseweg b4 2333 AL
Leiden, The Netherlands (R.B., C.S.G.A., P.J.J.H., R.O.)
The plant hormone auxin is a central regulator of plant development. In
Arabidopsis, the PINOID (PID) protein serine/threonine kinase is a key
component in the signaling of this phytohormone. To further investigate the
biological function of PID, we performed a screen for PID-interacting proteins
using the yeast two-hybrid system. Here, we show that PID interacts with two
calcium-binding proteins: TOUCH3 (TCH3), a calmodulin-related protein, and
PID-BINDING PROTEIN 1 (PBP1), a previously uncharacterized protein containing
putative EF-hand calcium-binding motifs. The interaction between PID and the
calcium-binding proteins is significant because it is calcium dependent and
requires an intact PID protein. Furthermore, the expression of all three genes
(PID, TCH3, and PBP1) is up-regulated by auxin. TCH3 and
PBP1 are not targets for phosphorylation by PID, suggesting that these
proteins act upstream of PID. PBP1 was found to stimulate the
autophosphorylation activity of PID, and calcium influx and calmodulin
inhibitors where found to enhance the activity of PID in vivo. Our results
indicate that TCH3 and PBP1 interact with the PID protein kinase and regulate
the activity of this protein in response to changes in calcium levels. This
work provides the first molecular evidence for the involvement of calcium in
auxin-regulated plant development.
Article, publication date, and citation information can be found at
www.plantphysiol.org/cgi/doi/10.1104/pp.103.019943.
1 Present address: Center of Applied Genetics, University of Agricultural
Sciences, Muthgasse 18, A–1190 Vienna, Austria.
*
Corresponding author; e-mail
offringa{at}rulbim.leidenuniv.nl;
fax 31–71–5274999.
Received January 6, 2003;
returned for revision February 4, 2003;
accepted April 4, 2003.
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