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Interactions among PIN-FORMED and P-Glycoprotein Auxin Transporters in Arabidopsis^[W]

Joshua J. Blakeslee^a,1,2, Anindita Bandyopadhyay^a,1, Ok Ran Lee^a,1,3, Jozef Mravec^b, Boosaree Titapiwatanakun^a, Michael Sauer^b, Srinivas N. Makam^a, Yan Cheng^a, Rodolphe Bouchard^c, Jií Adamec^d, Markus Geisler^c, Akitomo Nagashima^e, Tatsuya Sakai^e, Enrico Martinoia^c, Jií Friml^b, Wendy Ann Peer^a,4 and Angus S. Murphy^a

^a Department of Horticulture, Purdue University, West Lafayette, Indiana 47907-2010
^b Center for Plant Molecular Biology, University of Tübingen, D-72076 Tübingen, Germany
^c Institute of Plant Biology, Basel–Zurich Plant Science Center, University of Zurich, CH-8007 Zurich, Switzerland
^d Purdue Discovery Park, West Lafayette, Indiana 47907-2010
^e Genetic Regulatory Systems Research Team, RIKEN Plant Science Center, Kanagawa, 230-0045, Japan

⁴ To whom correspondence should be addressed. E-mail peerw{at}purdue.edu; fax 765-494-0391.

Directional transport of the phytohormone auxin is established primarily at the point of cellular efflux and is required for the establishment and maintenance of plant polarity. Studies in whole plants and heterologous systems indicate that PIN-FORMED (PIN) and P-glycoprotein (PGP) transport proteins mediate the cellular efflux of natural and synthetic auxins. However, aromatic anion transport resulting from PGP and PIN expression in nonplant systems was also found to lack the high level of substrate specificity seen in planta. Furthermore, previous reports that PGP19 stabilizes PIN1 on the plasma membrane suggested that PIN–PGP interactions might regulate polar auxin efflux. Here, we show that PGP1 and PGP19 colocalized with PIN1 in the shoot apex in Arabidopsis thaliana and with PIN1 and PIN2 in root tissues. Specific PGP–PIN interactions were seen in yeast two-hybrid and coimmunoprecipitation assays. PIN–PGP interactions appeared to enhance transport activity and, to a greater extent, substrate/inhibitor specificities when coexpressed in heterologous systems. By contrast, no interactions between PGPs and the AUXIN1 influx carrier were observed. Phenotypes of pin and pgp mutants suggest discrete functional roles in auxin transport, but pin pgp mutants exhibited phenotypes that are both additive and synergistic. These results suggest that PINs and PGPs characterize coordinated, independent auxin transport mechanisms but also function interactively in a tissue-specific manner.

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