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Cited by 1,530Plant Molecular Biology ResearchPlant nutrient uptake and metabolismPlant Parasitism and Resistance

Regulation of Polar Auxin Transport by AtPIN1 in Arabidopsis Vascular Tissue

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Gälweiler, Leo, Guan, Changhui, Müller, Andreas, Wisman, Ellen, Mendgen, Kurt, Yephremov, Alexander, Palme, Klaus·1998·Science
DOI 10.1126/science.282.5397.2226
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Abstract

Polar auxin transport controls multiple developmental processes in plants, including the formation of vascular tissue. Mutations affecting the PIN-FORMED (PIN1) gene diminish polar auxin transport in Arabidopsis thaliana inflorescence axes. The AtPIN1 gene was found to encode a 67-kilodalton protein with similarity to bacterial and eukaryotic carrier proteins, and the AtPIN1 protein was detected at the basal end of auxin transport–competent cells in vascular tissue. AtPIN1 may act as a transmembrane component of the auxin efflux carrier.

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Research works printed but not linked: 44

  1. G. Chang, R. Spencer, A. Lee, M. Barclay, D. Rees, data not shown. 30. A. Kreusch, P. J. Pfaffinger, C. F. Stevens, S. Choe, Nature 392, 945 (1998).
  2. X. R. Ou, P. Blount, R. J. Hoffman, C. Kung, Proc. Natl. Acad. Sci. U.S.A. 95, 11471 (1998).
  3. P. Blount, S. I. Sukharev, M. J. Schroeder, S. K. Nagle, C. Kung, ibid. 93, 11652 (1996).
  4. C. C. Hase, A. C. LeDain, B. Martinac, J. Membr. Biol. 157, 17 (1997).
  5. J. Biol. Chem. 270, 18329 (1995).
  6. H. A. Lester, Annu. Rev. Biophys. Biomol. Struct. 21, 267 (1992).
  7. N. Unwin, Nature 373, 37 (1995).
  8. J. Mol. Biol. 257, 586 (1996).
  9. S. Bailey, CCP4 Project, Acta Crystallogr. D 50, 760 (1994).
  10. Single-letter abbreviations for the amino acid resi- dues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr. 41. J. D. Thompson, T. J. Gibson, F. Plewniak, F. Jeanmou- gin, D. G. Higgins, Nucleic Acids Res. 25, 4876 (1997).
  11. R. M. Esnouf, J. Mol. Graphics 15, 132 (1997).
  12. P. J. Kraulis, J. Appl. Crystallogr. 24, 946 (1991).
  13. E. A. Merritt and D. J. Bacon, Methods Enzymol. 277, 505 (1997).
  14. B. Honig and A. Nicholls, Science 268, 1144 (1995).
  15. F. W. Went, Rec. Trav. Bot. Neerl. 25, 1 (1928);
  16. G. H. van der Weij, ibid. 31, 810 (1934).
  17. M. H. M. Goldsmith, Annu. Rev. Plant Physiol. 28, 439 (1977);
  18. T. L. Lomax, G. K. Muday, P. H. Rubery, in Plant Hormones, Physiology, Biochemistry and Molec- ular Biology, P. J. Davies, Ed. (Martinus Nijhoff, Klu- wer, Dordrecht, The Netherlands, 1995), pp. 509 - 530.
  19. T. Sachs, Pattern Formation in Plant Tissues (Cam- bridge Univ. Press, Cambridge, 1991);
  20. R. Aloni, in Plant Hormones, Physiology, Biochemistry and Molec- ular Biology, P. J. Davies, Ed. (Martinus Nijhoff, Klu- wer, Dordrecht, The Netherlands, 1995), pp. 531- 546.
  21. M. Jacobs and P. H. Rubery, Science 241, 346 (1988).
  22. M. Jacobs and S. F. Gilbert, ibid. 220, 1297 (1983).
  23. K. Okada, J. Ueda, M. K. Komaki, C. J. Bell, Y. Shimura, Plant Cell 3, 677 (1991).
  24. S. R. M. Bennett, J. Alvarez, G. Bossinger, D. R. Smyth, Plant J. 8, 505 (1995).
  25. G. H. Cardon, M. Frey, H. Saedler, A. Gierl, Plant J. 3, 773 (1993). The heterozygous Atpin1::En134 mutant (M 1 ), whose M 2 progeny segregated 3 :1 for wild-type and mutant phenotypes, was identified in generation S 6 [E. Wisman, G. H. Cardon, P. Fransz, H. Saedler, Plant Mol. Biol. 37, 989 (1998)]. Backcrosses of heterozygous Atpin1::En134 plants with wild type expressed the mu- tant phenotype only in the F 2 generation.
  26. Plant DNA sequences flanking the 5Ј end of En-1 in the Atpin1::En134 allele were cloned by a ligation- mediated PCR technique [P. R. Mueller and B. Wold, Science 246, 780 (1989);
  27. M. Frey, C. Stettner, A. Gierl, Plant J. 13, 717 (1998)] with En-1-and linker-specific oligonucleotides after Csp6 I restriction of genomic DNA and ligation of compatible linker DNA. The isolated flanking DNA was used as a probe to screen a cDNA library, prepared from suspension cells, for homologous clones that were then used to screen a genomic library of A. thaliana. The libraries were prepared from the ecotype Columbia and provided by the Arabidopsis DNA Centre, Cologne. Sequence analysis of the longest AtPIN1 cDNA (2276 base pairs) identified an open reading frame encoding 622 amino acids. An in-frame stop codon located up- stream to the first ATG suggested that the cDNA encodes a full-length protein. GenBank accession numbers are as follows: AF089084 (AtPIN1 cDNA) and AF089085 (AtPIN1 genomic DNA).
  28. F. Creusot et al., Plant J. 8, 763 (1995)]; provided by the Arabidopsis DNA Centre, Cologne} with a radiolabeled AtPIN1 probe, we identified a contig consisting of the over- lapping clones CIC6H1, CIC12G10, CIC12H9, and CIC9C4. Physical mapping was performed with the server http://cbil.humgen.upenn.edu/ϳatgc/physical- mapping.
  29. S. J. Singer, Annu. Rev. Cell Biol. 6, 247 (1990);
  30. M. D. Marger und M. H. Saier Jr., Trends Biochem. Sci. 18, 13 (1993);
  31. H. Logan, M. Basset, A.-A. Ve ´ry, H. Sente- nac, Physiol. Plant. 100, 1 (1997).
  32. C. Luschnig, R. A. Gaxiola, P. Grisafi, G. F. Fink, Genes Dev. 12, 2175 (1998);
  33. A. Mu ¨ller et al., EMBO J. 17, 6903 (1998). AtPIN2 (AF086906) and EIR1 (AF056026) were independently isolated and repre- sent the same genetic locus.
  34. To generate AtPIN1-specific polyclonal antibodies, we ligated a Rsa I fragment of the AtPIN1 cDNA encoding the antigenic peptide of AtPIN1 from amino acid 155 to 408 into the bacterial expression vector pQE-31 (Qiagen). This expression construct encoded a recombinant fusion protein with an NH 2 -terminal His 6 tag. After expression in Escherichia coli SG13009, the recombinant protein was affinity puri- fied on a Ni 2ϩ -nitrilotriacetic acid column as de- scribed by the Quiaexpressionist manual (Qiagen) and checked by SDS-polyacrylamide gel electro- phoresis [U. K. Laemmli, Nature 227, 680 (1970)].
  35. After immunization of rabbits (Eurogentec, Ougre ´e, Belgium), the polyclonal antiserum was affinity puri- fied against the recombinant AtPIN1 peptide [ J. Gu, G. Stephenson, M. J. Iadarola, Biotechniques 17, 257 (1994)] and diluted to a final protein concentration of 0.22 mg/ml. In protein immunoblot analysis the affinity-purified anti-AtPIN1 detected specifically the recombinant AtPIN1 peptide in bacterial extracts as well as a 67-kD protein in microsomal membrane fractions from A. thaliana [R. Zettl, J. Schell, K. Palme, Proc. Natl. Acad. Sci. U.S.A. 91, 689 (1994)].
  36. D. A. Morris and A. G. Thomas, J. Exp. Bot. 29, 147 (1978).
  37. A. Delbarre, P. Muller, V. Imhoff, J. Guern, Planta 198, 532 (1996).
  38. H. J. Klee, R. B. Horsch, M. A. Hinchee, M. B. Hein, N. L. Hoffmann, Genes Dev. 1, 86 (1987);
  39. C. Uggla, T. Moritz, G. Sandberg, B. Sundberg, Proc. Natl. Acad. Sci. U.S.A. 93, 9282 (1996).
  40. T. Teichmann et al., Eur. J. Biochem. 247, 826 (1997).
  41. F. M. Ausubel et al., Current Protocols in Molecular Biology (Green, Wiley, New York, 1993).
  42. Z. Schwarz-Sommer, A. Gierl, H. Cuypers, P. A. Peter- son, H. Saedler, EMBO J. 4, 579 (1985).
  43. Single-letter abbreviations for amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr. 27. J. Kyte and R. F. Doolittle, J. Mol. Biol. 157, 105 (1982).
  44. P. Chomczynski and N. Sacchi, Anal. Biochem. 162, 156 (1987). To check for equal RNA loading we rehybridized the Northern blots with ribosomal pro- tein large subunit 4 (RPL4) and ubiquitin carrier (UBC) probes. 29. Segments of inflorescence axes of 3-to 4-week-old A. thaliana ecotype Columbia (grown in a greenhouse at 18° to 24°C, with 16 hours of light) were fixed, paraffin embedded, and analyzed by in situ hybrid- ization as described (22), with the following modifi- cations. To generate AtPIN1-specific RNA probes, we inserted the Bgl II-Hind III fragment of the AtPIN1 cDNA (base pairs 602 to 1099) into the Bam HI-, Hind III-cleaved vector pBluescript SK-(Stratagene), J. P. Schwarz and J. E. Faller, JILA, University of Colo- rado and National Institute of Standards and Tech- nology, Boulder, CO 80309 -0440, USA.

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Authors
Gälweiler, L., Guan, C., Müller, A., Wisman, E., Mendgen, K., Yephremov, A., & Palme, K.
Affiliations
  • L. Gälweiler, C. Guan, A. Müller, and K. Palme are at the Max-Delbrück-Laboratorium in der Max-Planck-Gesellschaft, Carl-von-Linné-Weg 10, D-50829 Köln, Germany. E. Wisman and A. Yephremov are at the Max-Planck-Institut für Züchtungsforschung, Abteilung Molekulare Pflanzengenetik, Carl-von-Linné-Weg 10, D-50829 Köln, Germany. K. Mendgen is at the Universität Konstanz, Fakultät für Biologie/Phytopathologie, D-78457 Konstanz, Germany.
  • Max Planck Society (DE)
Corresponding author
Palme, Klaus
ORCID
Venue
Science
Publisher
American Association for the Advancement of Science (AAAS)
Published
1998-12-18
Volume
282
Issue
5397
Pages
2226-2230
ISSN
  • 0036-8075 (ISSN-L)
  • 1095-9203
DOI
10.1126/science.282.5397.2226
OpenAlex
W2038818582
Cited by
1,530
References
1
Citations by year
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FWCI
57.01
Type
journal article
Language
en
Open access
Licence
cc-by-nc-nd
Locations
  • Science (publishedVersion)
  • PubMed (publishedVersion)
  • KOPS (University of Konstanz) (submittedVersion, cc-by-nc-nd)
Funders
  • U.S. Department of Energy
  • Amgen
  • National Institutes of Health
Topics
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Molecular Biology Research
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant nutrient uptake and metabolism
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Parasitism and Resistance
Keywords
polar auxin transport, PIN-FORMED proteins, auxin efflux carriers, vascular tissue development, Arabidopsis thaliana, auxin transport
MeSH terms
Amino Acid Sequence, Biological Transport, Cloning, Molecular, DNA Transposable Elements, Indoleacetic Acids, Membrane Proteins, Molecular Sequence Data, Phenotype, Phthalimides, Blotting, Northern, Mutagenesis, Insertional, Genes, Plant, Arabidopsis, Plant Roots, Plant Stems, Proton-Motive Force, Contig Mapping, Membrane Transport Proteins, Arabidopsis Proteins

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