Allelic analyses of the Arabidopsis YUC1 locus reveal residues and domains essential for the functions of YUC family of flavin monooxygenases.
Allelic analyses of the Arabidopsis YUC1 locus reveal residues and domains essential for the functions of YUC family of flavin monooxygenases.
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DOI：
10.1111/j.1744-7909.2010.01007.x
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发表时间：
2011-01
影响因子：
11.4
通讯作者：
Zhao Y
中科院分区：
文献类型：
--
作者：
Hou X;Liu S;Pierri F;Dai X;Qu LJ;Zhao Y
黄素单加氧酶(FMOS)通过合成生长素和其他信号分子在植物生长发育中发挥重要作用。然而，植物FMO内部的结构和功能关系尚不清楚。在这里，我们定义了拟南芥YUC1FMO的重要残基和结构域，它是生长素生物合成的关键酶。我们先前已经证明，同时失活YUC1及其同源物YUC4会导致维管和花发育的严重缺陷。我们对yuc4突变体进行了诱变，并筛选出表型与yuc1、yuc4双突变体相似的突变体。在分离出的突变株中，有五个含有YUC1基因突变。有趣的是，在新的yuc1等位基因中发现的突变集中在两个在植物Fmo中高度保守的GXGXXG基序中。其中一个基序可能与黄素腺嘌呤二核苷酸(FAD)辅因子结合，另一个基序与烟酰胺腺嘌呤二核苷酸磷酸(NADPH)结合。我们还在yuc1中发现了Ser139到Phe的转换，这是一个位于两个核苷酸结合位点之间的突变。通过对一系列yuc1突变体的分析，我们确定了yuc1 FMO功能所必需的关键残基和基序。
Flavin monooxygenases (FMOs) play critical roles in plant growth and development by synthesizing auxin and other signaling molecules. However, the structure and function relationship within plant FMOs is not understood. Here we defined the important residues and domains of the Arabidopsis YUC1 FMO, a key enzyme in auxin biosynthesis. We previously showed that simultaneous inactivation of YUC1 and its homologue YUC4 caused severe defects in vascular and floral development. We mutagenized the yuc4 mutant and screened for mutants with phenotypes similar to those of yuc1 yuc4 double mutants. Among the isolated mutants, five of them contained mutations in the YUC1 gene. Interestingly, the mutations identified in the new yuc1 alleles were concentrated in the two GXGXXG motifs that are highly conserved among the plant FMOs. One such motif presumably binds to flavin adenine dinucleotide (FAD) cofactor and the other binds to nicotinamide adenine dinucleotide phosphate (NADPH). We also identified the Ser139 to Phe conversion in yuc1, a mutation that is located between the two nucleotide-binding sites. By analyzing a series of yuc1 mutants, we identified key residues and motifs essential for the functions of YUC1 FMO.
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