Arabidopsis AUX1 gene: A permease-like regulator of root gravitropism

Science

Volumn 273, Issue 5277, 1996, Pages 948-950

  Bennett, Malcolm J ^a   Marchant, Alan ^a   Green, Haydn G ^a   May, Sean T ^a   Ward, Sally P ^a   Millner, Paul A ^b   Walker, Amanda R ^c   Schulz, Burkhard ^d,e   Feldmann, Kenneth A ^d  

^a UNIVERSITY OF WARWICK   (United Kingdom)

^b UNIVERSITY OF LEEDS   (United Kingdom)

^c UNIVERSITY OF CAMBRIDGE   (United Kingdom)

^d UNIVERSITY OF ARIZONA   (United States)

^e UNIVERSITY OF COLOGNE   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

AUXIN; INDOLEACETIC ACID; PERMEASE; TRYPTOPHAN;

AMINO ACID SEQUENCE; ARTICLE; GENE MUTATION; GROWTH REGULATION; MOLECULAR CLONING; PLANT GROWTH; PLANT ROOT; PRIORITY JOURNAL; REGULATORY MECHANISM; SEQUENCE HOMOLOGY;

ARABIDOPSIS; EMBRYOPHYTA;

EID: 0029811396     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5277.948     Document Type: Article

References (48)

1. M. Estelle, Bioessays 14, 439 (1992).
2. J. Normanly, J. R. Cohen, G. R. Fink, Proc. Natl. Acad. Sci. U.S.A 90, 10355 (1993).
3. P. H. Rubery and A. R. Sheldrake, Planta 118, 101 (1974).
4. D. L. Rayle and R. E. Cleland, Plant Physiol. 99, 1271 (1992).
5. A. M. Jones, Annu. Rev. Plant Physiol. Plant Mol. Biol. 45, 394 (1994).
6. H. M. O. Leyser et al., Nature 364, 161 (1993).
7. M. L. Evans, Plant Physiol. 95, 1 (1991).
8. K. A. Feldmann, Plant J. 1, 71 (1991).
9. E. P. Maher and S. J. B. Martindale, Biochem. Genet. 18, 1041 (1980).
10. F. B. Pickett, A. K. Wilson, M. Estelle, Plant Physiol. 94, 1462 (1990).
11. Root gravitropic curvature was measured in 30 to 40 4-day-old seedlings from each of the 6000 Arizona T-DNA lines as described [B. L. Bullen et al., Plant Physiol. 93, 525 (1990)]. One agravitropic mutant, aux1-100, was identified as being allelic with aux1 when crossed with the aux1-7 mutant (10).
12. Root thigmotropism was measured as described [K. Okada and Y. Shimura, Science 250, 274 (1990)].
13. 8M [ecotype Wassilewskija (WS)].
14. To test for linkage between the T-DNA and aux1-100 phenotype, we selfed 77 agravitropic progeny from a plant heterozygous for aux1-100 and scored their progeny for kanamycin resistance. All of the progeny were kanamycin resistant, confirming that their agravitropic parents were homozygous for the T-DNA insert.
15. A 2.2-kbp fragment of plant-flanking DNA was plasmid rescued from aux1-100 genomic DNA [as described in (16)]. Southern hybridization experiments with this radiolabeled fragment highlighted a Hind III RFLP between the wild-type (ecotype WS) and aux1-100 DNA.
16. B. Schulz, M. J. Bennett, B. P. Dilkes, K. A. Feldmann, Plant Molecular Biology Manual K3 (Kluwer, Netherlands, 1994).
17. An Arabidopsis (ecotype WS) genomic library [as described in (76)] was screened with the radiolabeled 2.2-kbp fragment at high stringency [0.1 × standard saline citrate (SSC) and 0.1% SDS at 65°C] to isolate the homologous clone 20-1-1.
18. Mapping experiments with a recombinant inbred population [C. Lister and C. Dean, Plant J. 4, 745 (1993)] concluded that a Dra I RFLP encoded by 20-1-1 was located on chromosome 2, close to the AUX1 gene.
19. The PRL2 cDNA library [T. Newman et al., Plant Physiol. 106, 1241 (1994)] was screened at high stringency (0.1 × SSC and 0.1% SDS at 65°C) with a 20-1-1 probe, and multiple cDNA clones were characterized. A 5′ RACE (rapid amplification of cDNA ends) approach was used to confirm that the longest cDNA clone was full length.
20. EMBL accession number X98772.
21. Each aux1 allele was isolated as three overlapping polymerase chain reaction fragments and then sequenced directly as described [S. Khorana, R. F. Gagel, G. J. Cote, Nucl. Acids Res. 22, 3425 (1994)].
22. A 7.5-kbp Xho I-Bam HI fragment and a 4.4-kbp Eco RV-Bam HI fragment (Fig. 1) were subcloned into a Bin19-based kanamycin-resistant plant-transformation vector [M. Bevan, Nucl. Acids Res. 12, 8711 (1984)]. The XB7.5 and EB4.4 constructs were transformed into the aux1-7 mutant through use of a vacuum infiltration procedure [N. Bechtold et al., C. R. Acad. Sci. (Paris) 316, 1194 (1993)].
23. A 7.5-kbp Xho I-Bam HI fragment and a 4.4-kbp Eco RV-Bam HI fragment (Fig. 1) were subcloned into a Bin19-based kanamycin-resistant plant-transformation vector [M. Bevan, Nucl. Acids Res. 12, 8711 (1984)]. The XB7.5 and EB4.4 constructs were transformed into the aux1-7 mutant through use of a vacuum infiltration procedure [N. Bechtold et al., C. R. Acad. Sci. (Paris) 316, 1194 (1993)].
24. Restoration of auxin-sensitive root growth (12) and of gravitropic (10) and thigmotropic (11) bending responses were observed within the progeny of six independent XB7.5 and EB4.4 transformants.
25. Database searches were performed with the BLASTP program [S. F. Altschul et al., J. Mol. Biol. 215, 403 (1990)] at the National Center tor Bioinformatics. Alignments were performed with the gap algorithm.
26. W. B. Frommer, S. Hummel, J. W. Reismeier, Proc. Natl. Acad. Sci. U.S.A. 90, 5944 (1993); L. C. Hsu, T. J. Chiou, L. Chen, D. R. Bush, ibid., p. 7441; M. Kwart, B. Hirner, S. Hummel, W. B. Frommer, Plant J. 4, 993 (1993); W. N. Fischer, EMBL submissions S51168, S51169, and S51170; E. B. Lelanne, F. Vedel, R. DePaepe, EMBL submission U31932; D. Dillon and D. Stadler, Genetics 138, 61 (1994).
27. W. B. Frommer, S. Hummel, J. W. Reismeier, Proc. Natl. Acad. Sci. U.S.A. 90, 5944 (1993); L. C. Hsu, T. J. Chiou, L. Chen, D. R. Bush, ibid., p. 7441; M. Kwart, B. Hirner, S. Hummel, W. B. Frommer, Plant J. 4, 993 (1993); W. N. Fischer, EMBL submissions S51168, S51169, and S51170; E. B. Lelanne, F. Vedel, R. DePaepe, EMBL submission U31932; D. Dillon and D. Stadler, Genetics 138, 61 (1994).
28. W. B. Frommer, S. Hummel, J. W. Reismeier, Proc. Natl. Acad. Sci. U.S.A. 90, 5944 (1993); L. C. Hsu, T. J. Chiou, L. Chen, D. R. Bush, ibid., p. 7441; M. Kwart, B. Hirner, S. Hummel, W. B. Frommer, Plant J. 4, 993 (1993); W. N. Fischer, EMBL submissions S51168, S51169, and S51170; E. B. Lelanne, F. Vedel, R. DePaepe, EMBL submission U31932; D. Dillon and D. Stadler, Genetics 138, 61 (1994).
29. W. B. Frommer, S. Hummel, J. W. Reismeier, Proc. Natl. Acad. Sci. U.S.A. 90, 5944 (1993); L. C. Hsu, T. J. Chiou, L. Chen, D. R. Bush, ibid., p. 7441; M. Kwart, B. Hirner, S. Hummel, W. B. Frommer, Plant J. 4, 993 (1993); W. N. Fischer, EMBL submissions S51168, S51169, and S51170; E. B. Lelanne, F. Vedel, R. DePaepe, EMBL submission U31932; D. Dillon and D. Stadler, Genetics 138, 61 (1994).
30. W. B. Frommer, S. Hummel, J. W. Reismeier, Proc. Natl. Acad. Sci. U.S.A. 90, 5944 (1993); L. C. Hsu, T. J. Chiou, L. Chen, D. R. Bush, ibid., p. 7441; M. Kwart, B. Hirner, S. Hummel, W. B. Frommer, Plant J. 4, 993 (1993); W. N. Fischer, EMBL submissions S51168, S51169, and S51170; E. B. Lelanne, F. Vedel, R. DePaepe, EMBL submission U31932; D. Dillon and D. Stadler, Genetics 138, 61 (1994).
31. W. B. Frommer, S. Hummel, J. W. Reismeier, Proc. Natl. Acad. Sci. U.S.A. 90, 5944 (1993); L. C. Hsu, T. J. Chiou, L. Chen, D. R. Bush, ibid., p. 7441; M. Kwart, B. Hirner, S. Hummel, W. B. Frommer, Plant J. 4, 993 (1993); W. N. Fischer, EMBL submissions S51168, S51169, and S51170; E. B. Lelanne, F. Vedel, R. DePaepe, EMBL submission U31932; D. Dillon and D. Stadler, Genetics 138, 61 (1994).
32. AUX1 transmembrane predictions were performed with software described by the following researchers; B. Rost et al., Protein Sci. 4, 521 (1994); M. Claros and G. von Heijne, CABIOS 10, 685 (1994); D. M. Engelman, T. A. Steitz, A. Goldman, Annu. Rev. Biophys. Biophys. Chem. 15, 321 (1986).
33. AUX1 transmembrane predictions were performed with software described by the following researchers; B. Rost et al., Protein Sci. 4, 521 (1994); M. Claros and G. von Heijne, CABIOS 10, 685 (1994); D. M. Engelman, T. A. Steitz, A. Goldman, Annu. Rev. Biophys. Biophys. Chem. 15, 321 (1986).
34. AUX1 transmembrane predictions were performed with software described by the following researchers; B. Rost et al., Protein Sci. 4, 521 (1994); M. Claros and G. von Heijne, CABIOS 10, 685 (1994); D. M. Engelman, T. A. Steitz, A. Goldman, Annu. Rev. Biophys. Biophys. Chem. 15, 321 (1986).
35. T. L. Lomax, G. K. Muday, P. Rubery, in Plant Hormones and Their Role in Plant Growth and Development, P. J. Davies, Ed. (Martinus Nijhoft, Dordrecht, ed. 2, 1994), pp 509-530.
36. D. R. Bush, Annu. Rev. Plant Physiol. Plant Mol. Biol. 44, 513 (1993).
37. T. L. Lomax and G. R. Hicks, Biochem. Soc. Trans. 20, 64 (1992); R. Zettl et al., Proc. Natl. Acad. Sci. U.S.A 89, 480 (1992).
38. T. L. Lomax and G. R. Hicks, Biochem. Soc. Trans. 20, 64 (1992); R. Zettl et al., Proc. Natl. Acad. Sci. U.S.A 89, 480 (1992).
39. G. K. Muday and P. Haworth, Plant Physiol. Biochem. 32, 193 (1994).
40. S. Tsurumi and Y. Ohwaki, Plant Cell Physiol. 19, 1195 (1978).
41. K. H. Hasenstein and M. L. Evans, Plant Physiol. 86, 890 (1988).
42. A 350-bp fragment from the 5' end of the AUX1 cDNA was subcloned into the bluescript plasmid (Stratagene) and linearized with either Spe1 or Eco R1 in order to synthesize sense or antisense strand-specific AUX1 RNA probes, respectively. The RNA probes were labeled with digoxygenin during in vitro transcription (Boehringer Mannheim) and then hydrolyzed to approximately 150 bases in length and then quantified; 10 ng of each probe were hybridized with 2-day-old Arabidopsis seedlings through use of a whole-mount in situ procedure [as described by D. Ludevid, H. Hotte, E. Himelblau, M. J. Chrispeels, Plant Physiol. 100, 1633 (1992)].
43. S. Ward, A. Marchant, M. J. Bennett, unpublished data.
44. M. J. Vesper and C. L. Kuss, Planta 182, 486 (1990).
45. M. L. Evans, H. Ishikawa, M. Estelle, ibid. 194, 215 (1994).
46. S. T. May, A. Marchant, M. J. Bennett, unpublished data.
47. Abbreviations for the 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.
48. We thank S. C. Bennett, B. P. Dikes, M. Schweigert, W. Woodward, and N. Forsthoefel for technical assistance We thank M. Estelle for seeds of aux1-7, G. Roman and J. Ecker for seeds of aux1-21 and aux1-22, N. Olszewski for the pOCA18 cosmid vector, and the Arabidopsis Biological Resource Center at Ohio State University for supplying the PRL2 cDNA library. Special thanks to M. Estelle, J, Digby, R. Firn, and G. Scherer for useful discussion. Supported by NATO (to M.J.B.), the Agricultural Food Research Council Plant Molecular Biology II/Biotechnology and Biological Sciences Research Council (to A.M., H.G.G., S.P.W., S.T.M., P.A.M., and A.R.W.), Deutsche Forschungsgemeinschaft (to B.S.), and NSF (to K.A.F.).