Dumas B, Borel C, Herbert C, Maury J, Jacquet C, Balsse R, Esquerré-Tugayé M T
UMR 5546 CNRS-Université Paul Sabatier, Pôle de Biotechnologie Végétale, 24 Chemin de Borde Rouge, BP17 Auzeville, 31326 Castanet-Tolosan, France.
Gene. 2001 Jul 11;272(1-2):219-25. doi: 10.1016/s0378-1119(01)00536-4.
The gene CLPT1 (Colletotrichum lindemuthianum Protein Transport 1) encoding a Rab/GTPase was isolated from the filamentous fungus Colletotrichum lindemuthianum, the causal agent of bean anthracnose. At the amino acid level, CLPT1 shows between 54 and 80% identity to SEC4-like proteins, a class of molecules required for intracellular vesicular transport in yeasts. In particular, typical SEC4 domains involved in nucleotide binding and membrane attachment are present in the CLPT1 sequence. Functional identity of CLPT1 with SEC4 was confirmed by complementation of the Saccharomyces cerevisiae sec4-8 mutation. This is the first report of a gene involved in the control of intracellular vesicular trafficking in a phytopathogenic fungus. RNA blot analyses of CLPT1 expression were performed during in vitro growth of the fungus on synthetic media containing glucose or pectin, as single carbon source. The accumulation of CLPT1 mRNA was strongly increased on pectin, a plant cell wall polysaccharide that induces the production of extracellular pectinases, whereas the level of CLPT1 mRNA was below the detection threshold on glucose. These results suggest that CLPT1 is mainly involved in protein secretion and that the production of extracellular enzymes potentially involved in pathogenesis in filamentous fungi is sustained by induction of the genes involved in the secretory machinery.
从引起菜豆炭疽病的丝状真菌菜豆炭疽菌(Colletotrichum lindemuthianum)中分离出了编码Rab / GTPase的基因CLPT1(菜豆炭疽菌蛋白转运1)。在氨基酸水平上,CLPT1与SEC4样蛋白具有54%至80%的同一性,SEC4样蛋白是酵母细胞内囊泡运输所需的一类分子。特别是,CLPT1序列中存在参与核苷酸结合和膜附着的典型SEC4结构域。通过互补酿酒酵母sec4 - 8突变证实了CLPT1与SEC4的功能同一性。这是关于植物病原真菌中参与细胞内囊泡运输控制的基因的首次报道。在真菌于含有葡萄糖或果胶作为单一碳源的合成培养基上进行体外生长期间,对CLPT1表达进行了RNA印迹分析。在果胶(一种诱导细胞外果胶酶产生的植物细胞壁多糖)上,CLPT1 mRNA的积累显著增加,而在葡萄糖上CLPT1 mRNA的水平低于检测阈值。这些结果表明,CLPT1主要参与蛋白质分泌,并且丝状真菌中潜在参与致病过程的细胞外酶的产生是由分泌机制相关基因的诱导所维持的。