Osanai Takashi, Kuwahara Ayuko, Otsuki Hitomi, Saito Kazuki, Yokota Hirai Masami
RIKEN Center for Sustainable Resource Science, Suehiro-cho, Tsurumi-ku, Yokohama, Japan.
PRESTO, Japan Science and Technology Agency (JST), Honcho, Kawaguchi, Saitama, Japan.
Plant Cell Physiol. 2017 Apr 1;58(4):650-657. doi: 10.1093/pcp/pcx033.
Glutamine synthetase (GS) is an important enzyme for nitrogen assimilation, and GS2, encoded by GLN2, is the only plastid-type GS in Arabidopsis thaliana. A co-expression analysis suggested that the expression level of the gene encoding a uridylyltransferase-like protein, ACR11, is strongly correlated with GLN2 expression levels. Here we showed that the recombinant ACR11 protein increased GS2 activity in vitro by reducing the Km values of its substrate glutamine. A T-DNA insertion mutant of ACR11 exhibited a reduced GS activity under low nitrate conditions and reduced glutamine levels. Biochemical analyses revealed that ACR11 and GS2 interacted both in vitro and in vivo. These data demonstrate that ACR11 is an activator of GS2, giving it a mechanistic role in the nitrogen assimilation of A. thaliana.
谷氨酰胺合成酶(GS)是氮同化的一种重要酶,由GLN2编码的GS2是拟南芥中唯一的质体类型GS。一项共表达分析表明,编码类尿苷酰转移酶蛋白ACR11的基因的表达水平与GLN2的表达水平密切相关。在此我们表明,重组ACR11蛋白通过降低其底物谷氨酰胺的Km值在体外增加了GS2的活性。ACR11的T-DNA插入突变体在低硝酸盐条件下表现出GS活性降低和谷氨酰胺水平降低。生化分析表明,ACR11和GS2在体外和体内均相互作用。这些数据证明ACR11是GS2的激活剂,在拟南芥的氮同化中发挥机制作用。