Coordinated polar localization of auxin efflux carrier PIN1 by GNOM ARF GEF

Science

Volumn 286, Issue 5438, 1999, Pages 316-318

  Steinmann, Thomas ^a   Geldner, Niko ^a   Grebe, Markus ^a   Mangold, Stefan ^a   Jackson, Catherine L ^b   Paris, Sonia ^c   Gälweiler, Leo ^d   Palme, Klaus ^d   Jürgens, Gerd ^a  

^a UNIVERSITY OF TÜBINGEN   (Germany)

^b INSTITUT DE RECHERCHES INTERNATIONALES SERVIER   (France)

^c INSTITUT DE PHARMACOLOGIE MOLÉCULAIRE ET CELLULAIRE   (France)

^d Max Delbruck Laboratorium der Max Planck Gesellschaft   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

ADENOSINE DIPHOSPHATE RIBOSYLATION FACTOR; AUXIN; PHYTOHORMONE;

ARABIDOPSIS; ARTICLE; CELL POLARITY; EMBRYO DEVELOPMENT; NONHUMAN; PLANT ROOT; PRIORITY JOURNAL; PROTEIN BINDING; PROTEIN LOCALIZATION; PROTEIN TRANSPORT;

ADP-RIBOSYLATION FACTORS; ARABIDOPSIS; ARABIDOPSIS PROTEINS; BIOLOGICAL TRANSPORT; BREFELDIN A; CELL MEMBRANE; CELL POLARITY; CYTOSOL; GUANINE NUCLEOTIDE EXCHANGE FACTORS; INDOLEACETIC ACIDS; MEMBRANE PROTEINS; MEMBRANE TRANSPORT PROTEINS; PLANT ROOTS; PROTEIN CONFORMATION; RECOMBINANT PROTEINS; SEEDS; SOLUBILITY; SUBCELLULAR FRACTIONS;

ARABIDOPSIS;

EID: 0033536692     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.286.5438.316     Document Type: Article

References (32)

1. F. W. Went, Rec. Trav. Bot. Neerl. 25, 1 (1929).
2. T. Sachs, Development (suppl. 1), 83 (1991).
3. L. Gälweiler et al., Science 282, 2226 (1998).
4. T. Berleth and G. Jürgens, Development 118, 575 (1993).
5. G. K. Przemeck, J. Mattsson, C. S. Hardtke, Z. R. Sung, T. Berleth, Planta 200, 229 (1996); T. Ulmasov, G. Hagen, T. J. Guilfoyle, Science 276, 1865 (1997); C. S. Hardtke and T. Berleth, EMBO J. 17, 1405 (1998).
6. G. K. Przemeck, J. Mattsson, C. S. Hardtke, Z. R. Sung, T. Berleth, Planta 200, 229 (1996); T. Ulmasov, G. Hagen, T. J. Guilfoyle, Science 276, 1865 (1997); C. S. Hardtke and T. Berleth, EMBO J. 17, 1405 (1998).
7. G. K. Przemeck, J. Mattsson, C. S. Hardtke, Z. R. Sung, T. Berleth, Planta 200, 229 (1996); T. Ulmasov, G. Hagen, T. J. Guilfoyle, Science 276, 1865 (1997); C. S. Hardtke and T. Berleth, EMBO J. 17, 1405 (1998).
8. T. Hamann, U. Mayer, G. Jürgens, Development 126, 1387 (1999).
9. U. Mayer, G. Büttner and G. Jürgens, ibid. 117, 149 (1993).
10. C. W. Vroemen et al., Plant Cell 8, 783 (1996).
11. M. Busch, U. Mayer, G. Jürgens, Mol. Gen. Genet. 250, 681 (1996).
12. A. Peyroche, S. Paris, C. L. Jackson, Nature 384, 479 (1996).
13. Yeast strain CJY052-10-2/pgea1-19 (10) (permissive temperature 25°C, restrictive temperature 30°C) was transformed with pYX242 alone, and the rescue plasmid containing the entire open reading frame of GNOM amplified from plasmid c96 (9) and directionally ligated into yeast expression vector pYX242 (Invitrogen) by using 5′ Nco I and 3′ Mlu I sites.
14. M. Franco, P. Chardin, M. Chabre, S. Paris, J. Biol. Chem. 270, 1337 (1995).
15. 2-terminally His-tagged version of pYX242-GNOM was constructed by three-point ligation, in which a 5′ Eco RI, 3′ Nco I 6xHis adapter were inserted together with a 5′ Nco I, 3′ Mlu I-digested full-length GNOM fragment into an Eco RI, Mlu I-digested vector. Transformants of yeast strain EGY48 with pYX242-His-GNOM were grown at 30°C on -Leu yeast selective medium and proteins were purified as described (10). Cells were resuspended in 50 ml of buffer A plus proteinase inhibitors and lysed by grinding in liquid nitrogen in the presence of glass beads. Expressed proteins were purified by Ni-nitrilotriacetic acid affinity chromatography, and the control fraction from vector-transformed yeast strain contained the same contaminating bands as the GNOM fraction [assessed by silver staining (24)].
16. 2-terminal 6xHis-tag fusion protein in Escherichia coli (Qiaexpress, Qiagen) and purified for immunization of rabbits (15).
17. M. H. Lauber et al., J. Cell. Biol. 139, 1485 (1997).
18. The antiserum cross-reacted with other proteins. The same response was observed for an antiserum against the COOH-terminal region of GNOM (24). Attempts to purify the antisera by affinity chromatography and immunoadsorption did not remove the cross-reacting antibodies, and no distinct differences in immunolocalization of GNOM were observed between wild-type and gnom mutant cells (24).
19. +-ATPase (adenosine triphosphatase) (15) were found by immunolocalization to accumulate at their respective membrane compartments in wild-type and gnom embryos (24).
20. A. Peyroche et al., Mol. Cell 3, 275 (1999); J. Lippincott-Schwartz, L. C. Yuan, J. S. Bonifacino, R. D. Klausner, Cell 56, 801 (1989);
21. A. Peyroche et al., Mol. Cell 3, 275 (1999); J. Lippincott-Schwartz, L. C. Yuan, J. S. Bonifacino, R. D. Klausner, Cell 56, 801 (1989);
22. B. Satiat Jeunemaitre and C. Hawes, J. Cell Sci. 103, 1153 (1992).
23. J. M. Park et al., Plant Physiol. 115, 763 (1997).
24. Signals on protein immunoblots from four experiments were scanned with a Storm Phosphoimager (Molecular Dynamics) and quantitated with Imagequant.
25. A. Delbarre, P. Mueller, J. Guern, Plant. Physiol. 116, 833 (1998); D. A. Morris and J. S. Robinson, Planta 205, 606 (1998).
26. A. Delbarre, P. Mueller, J. Guern, Plant. Physiol. 116, 833 (1998); D. A. Morris and J. S. Robinson, Planta 205, 606 (1998).
27. Seven-day-old seedlings were incubated in liquid growth medium containing 100 μM BFA or an equal volume of dimethyl sulfoxide for 30 min or 2 hours, fixed for 30 min, and processed on gelatin-coated slides as described (15). Antibody incubation was for 4.5 hours with vacuum infiltration for 10 min at the beginning of each step. Photographs were taken with a Zeiss microscope (Axiophot).
28. K. Hadfi, V. Speth, G. Neuhaus, Development 125, 879 (1998).
29. T. Steinmann et al., data not shown.
30. G. Jürgens and U. Mayer, in EMBRYOS. Colour Altas of Development, J. B. L. Bard, Ed. (Wolfe, London, 1994), pp. 7-21.
31. Immunofluorescence localization in whole-mount preparations was done as described (15). Primary antibody anti-PIN1 was diluted 1:150, and Cy3-conjugated anti-rabbit secondary antibody (Dianova) was diluted 1:600. Confocal laser-scanning microscopy was done with the TCS-NT program (Leica). Standard scanning conditions: 100× objective, 1-to 1.5-fold zoomed.
32. We thank M. Kientz for technical assistance, H. Schwarz for immunizing rabbits, W. Michalke for providing the anti-PM ATPase monoclonal antibody, A. Peyroche for constructing the gea1-19 yeast strain, and T. Hamann, M. Heese, M. Hülskamp, T. Laux, and U. Mayer for critical reading. Funded by the Deutsche Forschungsgemeinschaft grant Ju179/3-4 and Leibniz Programm.