Structure-Function Analysis of the Presumptive Arabidopsis Auxin Permease AUX1[W]
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- Ranjan Swarup
- School of Biosciences, Sutton Bonington Campus, University of Nottingham, United Kingdom
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- Joanna Kargul
- School of Biosciences, Sutton Bonington Campus, University of Nottingham, United Kingdom
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- Alan Marchant
- School of Biosciences, Sutton Bonington Campus, University of Nottingham, United Kingdom
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- Daniel Zadik
- School of Biosciences, Sutton Bonington Campus, University of Nottingham, United Kingdom
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- Abidur Rahman
- Centre for Support to Research and Education Activities Isotope Division, Kobe University, Kobe, Japan
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- Rebecca Mills
- School of Biological Sciences, University of Southampton, United Kingdom
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- Anthony Yemm
- School of Biosciences, Sutton Bonington Campus, University of Nottingham, United Kingdom
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- Sean May
- School of Biosciences, Sutton Bonington Campus, University of Nottingham, United Kingdom
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- Lorraine Williams
- School of Biological Sciences, University of Southampton, United Kingdom
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- Paul Millner
- School of Biochemistry and Molecular Biology, University of Leeds, Leeds, United Kingdom
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- Seiji Tsurumi
- Centre for Support to Research and Education Activities Isotope Division, Kobe University, Kobe, Japan
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- Ian Moore
- Plant Sciences, University of Oxford, United Kingdom
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- Richard Napier
- Warwick-HRI, University of Warwick, Wellesbourne, United Kingdom
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- Ian D. Kerr
- School of Biomedical Sciences, Queens Medical Centre, University of Nottingham, United Kingdom
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- Malcolm J. Bennett
- School of Biosciences, Sutton Bonington Campus, University of Nottingham, United Kingdom
書誌事項
- 公開日
- 2004-11-01
- 権利情報
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- https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model
- DOI
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- 10.1105/tpc.104.024737
- 公開者
- Oxford University Press (OUP)
説明
<jats:title>Abstract</jats:title><jats:p>We have investigated the subcellular localization, the domain topology, and the amino acid residues that are critical for the function of the presumptive Arabidopsis thaliana auxin influx carrier AUX1. Biochemical fractionation experiments and confocal studies using an N-terminal yellow fluorescent protein (YFP) fusion observed that AUX1 colocalized with plasma membrane (PM) markers. Because of its PM localization, we were able to take advantage of the steep pH gradient that exists across the plant cell PM to investigate AUX1 topology using YFP as a pH-sensitive probe. The YFP-coding sequence was inserted in selected AUX1 hydrophilic loops to orient surface domains on either apoplastic or cytoplasmic faces of the PM based on the absence or presence of YFP fluorescence, respectively. We were able to demonstrate in conjunction with helix prediction programs that AUX1 represents a polytopic membrane protein composed of 11 transmembrane spanning domains. In parallel, a large aux1 allelic series containing null, partial-loss-of-function, and conditional mutations was characterized to identify the functionally important domains and amino acid residues within the AUX1 polypeptide. Whereas almost all partial-loss-of-function and null alleles cluster in the core permease region, the sole conditional allele aux1-7 modifies the function of the external C-terminal domain.</jats:p>
収録刊行物
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- The Plant Cell
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The Plant Cell 16 (11), 3069-3083, 2004-11-01
Oxford University Press (OUP)