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Plant Cell, Vol. 10, 231-244, Copyright © 1998, American Society of Plant Physiologists

The DWF4 Gene of Arabidopsis Encodes a Cytochrome P450 That Mediates Multiple 22{alpha}-Hydroxylation Steps in Brassinosteroid Biosynthesis

Sunghwa Choea, Brian P. Dilkesa, Shozo Fujiokab, Suguru Takatsutoc, Akira Sakuraib, and Kenneth A. Feldmanna
a Department of Plant Sciences, University of Arizona, Tucson, Arizona 85721
b Institute of Physical and Chemical Research (RIKEN), Wako-shi, Saitama 351-01, Japan
c Department of Chemistry, Joetsu University of Education, Joetsu-shi, Niigata 943, Japan

Correspondence to: Kenneth A. Feldmann, feldmann{at}ag.arizona.edu (E-mail), 520-621-7186 (fax).

dwarf4 (dwf4) mutants of Arabidopsis display a dwarfed phenotype due to a lack of cell elongation. Dwarfism could be rescued by the application of brassinolide, suggesting that DWF4 plays a role in brassinosteroid (BR) biosynthesis. The DWF4 locus is defined by four mutant alleles. One of these is the result of a T-DNA insertion. Plant DNA flanking the insertion site was cloned and used as a probe to isolate the entire DWF4 gene. Sequence analysis revealed that DWF4 encodes a cytochrome P450 monooxygenase with 43% identity to the putative Arabidopsis steroid hydroxylating enzyme CONSTITUTIVE PHOTOMORPHOGENESIS AND DWARFISM. Sequence analysis of two other mutant alleles revealed deletions or a premature stop codon, confirming that DWF4 had been cloned. This sequence similarity suggests that DWF4 functions in specific hydroxylation steps during BR biosynthesis. In fact, feeding studies utilizing BR intermediates showed that only 22{alpha}-hydroxylated BRs rescued the dwf4 phenotype, confirming that DWF4 acts as a 22{alpha}-hydroxylase.




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Plant Physiol.Home page
R. G. Winkler, M. R. Frank, D. W. Galbraith, R. Feyereisen, and K. A. Feldmann
Systematic Reverse Genetics of Transfer-DNA-Tagged Lines of Arabidopsis . Isolation of Mutations in the Cytochrome P450 Gene Superfamily
Plant Physiology, November 1, 1998; 118(3): 743 - 750.
[Abstract] [Full Text]


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Plant CellHome page
U. Klahre, T. Noguchi, S. Fujioka, S. Takatsuto, T. Yokota, T. Nomura, S. Yoshida, and N.-H. Chua
The Arabidopsis DIMINUTO/DWARF1 Gene Encodes a Protein Involved in Steroid Synthesis
PLANT CELL, October 1, 1998; 10(10): 1677 - 1690.
[Abstract] [Full Text]


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Plant Physiol.Home page
H. Schaller, P. Bouvier-Navé, and P. Benveniste
Overexpression of an Arabidopsis cDNA Encoding a Sterol-C241-Methyltransferase in Tobacco Modifies the Ratio of 24-Methyl Cholesterol to Sitosterol and Is Associated with Growth Reduction
Plant Physiology, October 1, 1998; 118(2): 461 - 469.
[Abstract] [Full Text]


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Proc. Natl. Acad. Sci. USAHome page
C. A. Helliwell, C. C. Sheldon, M. R. Olive, A. R. Walker, J. A. D. Zeevaart, W. J. Peacock, and E. S. Dennis
Cloning of the Arabidopsis ent-kaurene oxidase gene GA3
PNAS, July 21, 1998; 95(15): 9019 - 9024.
[Abstract] [Full Text] [PDF]


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Plant CellHome page
R. Azpiroz, Y. Wu, J. C. LoCascio, and K. A. Feldmann
An Arabidopsis Brassinosteroid-Dependent Mutant Is Blocked in Cell Elongation
PLANT CELL, February 1, 1998; 10(2): 219 - 230.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
T. Asami, M. Mizutani, S. Fujioka, H. Goda, Y. K. Min, Y. Shimada, T. Nakano, S. Takatsuto, T. Matsuyama, N. Nagata, et al.
Selective Interaction of Triazole Derivatives with DWF4, a Cytochrome P450 Monooxygenase of the Brassinosteroid Biosynthetic Pathway, Correlates with Brassinosteroid Deficiency in Planta
J. Biol. Chem., July 6, 2001; 276(28): 25687 - 25691.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
C. A. Helliwell, P. M. Chandler, A. Poole, E. S. Dennis, and W. J. Peacock
The CYP88A cytochrome P450, ent-kaurenoic acid oxidase, catalyzes three steps of the gibberellin biosynthesis pathway
PNAS, February 13, 2001; 98(4): 2065 - 2070.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
J. Li, K. A. Lease, F. E. Tax, and J. C. Walker
BRS1, a serine carboxypeptidase, regulates BRI1 signaling in Arabidopsis thaliana
PNAS, May 8, 2001; 98(10): 5916 - 5921.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
I. Barlier, M. Kowalczyk, A. Marchant, K. Ljung, R. Bhalerao, M. Bennett, G. Sandberg, and C. Bellini
The SUR2 gene of Arabidopsis thaliana encodes the cytochrome P450 CYP83B1, a modulator of auxin homeostasis
PNAS, December 19, 2000; 97(26): 14819 - 14824.
[Abstract] [Full Text] [PDF]




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