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J. Biol. Chem., Vol. 281, Issue 41, 30603-30612, October 13, 2006

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Immunophilin-like TWISTED DWARF1 Modulates Auxin Efflux Activities of Arabidopsis P-glycoproteins^*

Rodolphe Bouchard, Aurélien Bailly, Joshua J. Blakeslee, Sophie C. Oehring, Vincent Vincenzetti, Ok Ran Lee¹, Ivan Paponov^¶, Klaus Palme^¶, Stefano Mancuso^||, Angus S. Murphy, Burkhard Schulz, and Markus Geisler²

From the Zurich-Basel Plant Science Center, University of Zurich, Institute of Plant Biology, and Molecular Plant Physiology, CH-8008 Zurich, Switzerland, the Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, Indiana 47907, the ^¶Institut für Biologie II, Universität Freiburg, D-79104 Freiburg, Germany, and the ^||Department of Horticulture, University of Firenze, I-50019 Sesto Fiorentino, Italy

The immunophilin-like protein TWISTED DWARF1 (TWD1/FKBP42) has been shown to physically interact with the multidrug resistance/P-glycoprotein (PGP) ATP-binding cassette transporters PGP1 and PGP19 (MDR1). Overlapping phenotypes of pgp1/pgp19 and twd1 mutant plants suggested a positive regulatory role of TWD1 in PGP-mediated export of the plant hormone auxin, which controls plant development. Here, we provide evidence at the cellular and plant levels that TWD1 controls PGP-mediated auxin transport. twd1 and pgp1/pgp19 cells showed greatly reduced export of the native auxin indole-3-acetic acid (IAA). Constitutive overexpression of PGP1 and PGP19, but not TWD1, enhanced auxin export. Coexpression of TWD1 and PGP1 in yeast and mammalian cells verified the specificity of the regulatory effect. Employing an IAA-specific microelectrode demonstrated that IAA influx in the root elongation zone was perturbed and apically shifted in pgp1/pgp19 and twd1 roots. Mature roots of pgp1/pgp19 and twd1 plants revealed elevated levels of free IAA, which seemed to account for agravitropic root behavior. Our data suggest a novel mode of PGP regulation via FK506-binding protein-like immunophilins, implicating possible alternative strategies to overcome multidrug resistance.

Received for publication, May 12, 2006 , and in revised form, July 18, 2006.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1–6.

¹ Present address: Department of Biology Chungnam National University, Daejeon 305–764, Korea.

² To whom correspondence should be addressed: Inst. of Plant Biology, University of Zurich, Zolliker Strasse 107, CH-8008 Zurich, Switzerland. Tel.: 41-1-634-8276; Fax: 41-1-634-8204; E-mail: markus.geisler{at}botinst.unizh.ch.

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