Goto Derek B, Ogi Makoto, Kijima Fumiko, Kumagai Tomoko, van Werven Folkert, Onouchi Hitoshi, Naito Satoshi
Laboratory of Molecular Biology, Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Japan.
Genes Genet Syst. 2002 Apr;77(2):89-95. doi: 10.1266/ggs.77.89.
Met-overaccumulating mutants provide a powerful genetic tool for examining both the regulation of the Met biosynthetic pathway and in vivo developmental responses of gene expression to altered Met levels. We have previously reported the identification of two Arabidopsis thaliana Met over-accumulation (mto) mutants, mto1-1 and mto2-1, that carry mutations in the genes encoding cystathionine gamma-synthase (CGS) and threonine synthase (TS), respectively. A third mutant, mto3-1, has recently been reported to carry a mutation in the gene encoding S-adenosylmethionine synthetase 3 (SAMS3). Here, we report the isolation of a new ethionine-resistant A. thaliana mutant that over-accumulates soluble Met approximately 20-fold in young rosettes. The causal mutation was determined to be a single, recessive mutation that was mapped to chromosome 3. Sequence analysis identified a single nucleotide change in the gene encoding SAMS3 that was distinct from the mto3-1 mutation and altered the amino acid sequence of the enzyme active site. This mutation was therefore referred to as mto3-2. Although Met over-accumulation in the mto3-2 mutant was similar to that in the mto2-1 mutant, CGS mRNA levels did not respond to the mto3-2 mutation and were similar to that in equivalent wild-type plants.
甲硫氨酸过度积累突变体为研究甲硫氨酸生物合成途径的调控以及基因表达对体内甲硫氨酸水平变化的发育反应提供了强大的遗传工具。我们之前报道过鉴定出两个拟南芥甲硫氨酸过度积累(mto)突变体,mto1-1和mto2-1,它们分别在编码胱硫醚γ-合酶(CGS)和苏氨酸合酶(TS)的基因中携带突变。最近有报道称,第三个突变体mto3-1在编码S-腺苷甲硫氨酸合成酶3(SAMS3)的基因中携带突变。在此,我们报告分离出一个新的抗乙硫氨酸的拟南芥突变体,该突变体在幼嫩莲座叶中可溶性甲硫氨酸过度积累约20倍。经确定,其致病突变是一个单隐性突变,定位于3号染色体。序列分析在编码SAMS3的基因中鉴定出一个单核苷酸变化,该变化与mto3-1突变不同,并改变了该酶活性位点的氨基酸序列。因此,这个突变被称为mto3-2。虽然mto3-2突变体中甲硫氨酸的过度积累与mto2-1突变体相似,但CGS mRNA水平对mto3-2突变没有反应,与相应野生型植物中的水平相似。