Enquiry into the topology of plasma membrane localized PIN auxin transport components

Nodzyński, Tomasz and Vanneste, Steffen and Zwiewka, Marta and Pernisová, Markéta and Hejátko, Jan and Friml, Jiří (2016) Enquiry into the topology of plasma membrane localized PIN auxin transport components. Molecular Plant, 9 (11). pp. 1504-1519. ISSN 1752-9867

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Official URL: http://dx.doi.org/10.1016/j.molp.2016.08.010


Auxin directs plant ontogenesis via differential accumulation within tissues depending largely on the activity of PIN proteins that mediate auxin efflux from cells and its directional cell-to-cell transport. Regardless of the developmental importance of PINs, the structure of these transporters is poorly characterized. Here, we present experimental data concerning protein topology of plasma membrane-localized PINs. Utilizing approaches based on pH-dependent quenching of fluorescent reporters combined with immunolocalization techniques, we mapped the membrane topology of PINs and further cross-validated our results using available topology modeling software. We delineated the topology of PIN1 with two transmembrane (TM) bundles of five α-helices linked by a large intracellular loop and a C-terminus positioned outside the cytoplasm. Using constraints derived from our experimental data, we also provide an updated position of helical regions generating a verisimilitude model of PIN1. Since the canonical long PINs show a high degree of conservation in TM domains and auxin transport capacity has been demonstrated for Arabidopsis representatives of this group, this empirically enhanced topological model of PIN1 will be an important starting point for further studies on PIN structure-function relationships. In addition, we have established protocols that can be used to probe the topology of other plasma membrane proteins in plants. © 2016 The Authors

Item Type: Article
DOI: 10.1016/j.molp.2016.08.010
Uncontrolled Keywords: Topology, arabidopsis thaliana, auxin efflux carriers, plasma membrane protein
Subjects: 500 Science > 580 Plants > 581 Botany
Research Group: Friml Group
SWORD Depositor: Sword Import User
Depositing User: Sword Import User
Date Deposited: 26 Jan 2017 14:20
Last Modified: 05 Sep 2017 09:32
URI: https://repository.ist.ac.at/id/eprint/746

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