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The GA2 Locus of Arabidopsis thaliana Encodes ent-Kaurene Synthase of Gibberellin Biosynthesis

Shinjiro Yamaguchi*, Tai-ping Sun, Hiroshi Kawaide, and Yuji Kamiya

Frontier Research Program, The Institute of Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako-shi, Saitama 351-01, Japan (S.Y., H.K., Y.K.); and Developmental, Cell and Molecular Biology Group, Department of Botany, Box 91000, Duke University, Durham, North Carolina 27708-1000 (S.Y., T.-p.S.)

The ga2 mutant of Arabidopsis thaliana is a gibberellin-deficient dwarf. Previous biochemical studies have suggested that the ga2 mutant is impaired in the conversion of copalyl diphosphate to ent-kaurene, which is catalyzed by ent-kaurene synthase (KS). Overexpression of the previously isolated KS cDNA from pumpkin (Cucurbita maxima) (CmKS) in the ga2 mutant was able to complement the mutant phenotype. A genomic clone coding for KS, AtKS, was isolated from A. thaliana using CmKS cDNA as a heterologous probe. The corresponding A. thaliana cDNA was isolated and expressed in Escherichia coli as a fusion protein. The fusion protein showed enzymatic activity that converted [3H]copalyl diphosphate to [3H]ent-kaurene. The recombinant AtKS protein derived from the ga2-1 mutant is truncated by 14 kD at the C-terminal end and does not contain significant KS activity in vitro. Sequence analysis revealed that a C-2099 to T base substitution, which converts Gln-678 codon to a stop codon, is present in the AtKS cDNA from the ga2-1 mutant. Taken together, our results show that the GA2 locus encodes KS.

*   Corresponding author; e-mail shinjiro{at}acpub.duke.edu; fax 1-919-613-8177.

Plant Physiol. (1998) 116: 1271-1278
Copyright Clearance Center:   0032-0889/98/116/1271/08
© 1998 American Society of Plant Physiologists




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