Hong Z, Zhang Z, Olson J M, Verma D P
Department of Molecular Genetics and Plant Biotechnology Center, Ohio State University, 1060 Carmack Road, Columbus, Ohio 43210-1002, USA.
Plant Cell. 2001 Apr;13(4):769-79. doi: 10.1105/tpc.13.4.769.
Using phragmoplastin as a bait, we isolated an Arabidopsis cDNA encoding a novel UDP-glucose transferase (UGT1). This interaction was confirmed by an in vitro protein--protein interaction assay using purified UGT1 and radiolabeled phragmoplastin. Protein gel blot results revealed that UGT1 is associated with the membrane fraction and copurified with the product-entrapped callose synthase complex. These data suggest that UGT1 may act as a subunit of callose synthase that uses UDP-glucose to synthesize callose, a 1,3-beta-glucan. UGT1 also interacted with Rop1, a Rho-like protein, and this interaction occurred only in its GTP-bound configuration, suggesting that the plant callose synthase may be regulated by Rop1 through the interaction with UGT1. The green fluorescent protein--UGT1 fusion protein was located on the forming cell plate during cytokinesis. We propose that UGT1 may transfer UDP-glucose from sucrose synthase to the callose synthase and thus help form a substrate channel for the synthesis of callose at the forming cell plate.
我们以成膜体素作为诱饵,分离出了拟南芥的一个编码新型尿苷二磷酸葡萄糖转移酶(UGT1)的cDNA。通过使用纯化的UGT1和放射性标记的成膜体素进行体外蛋白质-蛋白质相互作用试验,证实了这种相互作用。蛋白质凝胶印迹结果显示,UGT1与膜组分相关,并与包裹产物的胼胝质合酶复合物一起共纯化。这些数据表明,UGT1可能作为胼胝质合酶的一个亚基,利用尿苷二磷酸葡萄糖合成胼胝质(一种1,3-β-葡聚糖)。UGT1还与一种类Rho蛋白Rop1相互作用,且这种相互作用仅在其结合GTP的构象中发生,这表明植物胼胝质合酶可能通过与UGT1的相互作用受Rop1调控。绿色荧光蛋白-UGT1融合蛋白在胞质分裂期间定位于正在形成的细胞板上。我们推测,UGT1可能将尿苷二磷酸葡萄糖从蔗糖合酶转移至胼胝质合酶,从而有助于在正在形成的细胞板上形成用于胼胝质合成的底物通道。