Schomburg Fritz M, Bizzell Colleen M, Lee Dong Ju, Zeevaart Jan A D, Amasino Richard M
Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Plant Cell. 2003 Jan;15(1):151-63. doi: 10.1105/tpc.005975.
Degradation of active C(19)-gibberellins (GAs) by dioxygenases through 2beta-hydroxylation yields inactive GA products. We identified two genes in Arabidopsis (AtGA2ox7 and AtGA2ox8), using an activation-tagging mutant screen, that encode 2beta-hydroxylases. GA levels in both activation-tagged lines were reduced significantly, and the lines displayed dwarf phenotypes typical of mutants with a GA deficiency. Increased expression of either AtGA2ox7 or AtGA2ox8 also caused a dwarf phenotype in tobacco, indicating that the substrates for these enzymes are conserved. AtGA2ox7 and AtGA2ox8 are more similar to each other than to other proteins encoded in the Arabidopsis genome, indicating that they may constitute a separate class of GA-modifying enzymes. Indeed, enzymatic assays demonstrated that AtGA2ox7 and AtGA2ox8 both perform the same GA modification: 2beta-hydroxylation of C(20)-GAs but not of C(19)-GAs. Lines containing increased expression of AtGA2ox8 exhibited a GA dose-response curve for stem elongation similar to that of the biosynthetic mutant ga1-11. Double loss-of-function Atga2ox7 Atga2ox8 mutants had twofold to fourfold higher levels of active GAs and displayed phenotypes associated with excess GAs, such as early bolting in short days, resistance to the GA biosynthesis inhibitor ancymidol, and decreased mRNA levels of AtGA20ox1, a gene in the GA biosynthetic pathway.
双加氧酶通过2β-羟基化作用使活性C(19)-赤霉素(GAs)降解,产生无活性的GA产物。我们通过激活标签突变体筛选在拟南芥中鉴定出两个基因(AtGA2ox7和AtGA2ox8),它们编码2β-羟化酶。两个激活标签系中的GA水平均显著降低,且这些系表现出GA缺乏突变体典型的矮化表型。AtGA2ox7或AtGA2ox8的表达增加也导致烟草出现矮化表型,表明这些酶的底物是保守的。AtGA2ox7和AtGA2ox8彼此之间的相似性高于与拟南芥基因组中编码的其他蛋白质的相似性,这表明它们可能构成一类单独的GA修饰酶。实际上,酶活性测定表明AtGA2ox7和AtGA2ox8都进行相同的GA修饰:C(20)-GAs的2β-羟基化而非C(19)-GAs的2β-羟基化。AtGA2ox8表达增加的株系表现出与生物合成突变体ga1-11相似的茎伸长GA剂量反应曲线。Atga2ox7 Atga2ox8双功能缺失突变体的活性GA水平高出两到四倍,并表现出与GA过量相关的表型,如短日照下早抽薹、对GA生物合成抑制剂嘧啶醇的抗性以及GA生物合成途径中的基因AtGA20ox1的mRNA水平降低。