Guillén-Navarro Karina, Araíza Gisela, García-de los Santos Alejandro, Mora Yolanda, Dunn Michael F
Programa de Ingeniería Metabólica, Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, A. P. 565-A, Cuernavaca, Morelos, Mexico.
FEMS Microbiol Lett. 2005 Sep 15;250(2):209-19. doi: 10.1016/j.femsle.2005.07.020.
Because Rhizobium etli CE3 is normally dependent on an external source of biotin and lacks orthodox biotin biosynthesis genes, we undertook an analysis of biotin uptake in this organism. By complementation of a Sinorhizobium meliloti bioM mutant we isolated an R. etli chromosomal region encoding homologs of the S. meliloti bioMNB genes, whose products have been implicated in intracellular biotin retention in that organism. Disruption of the R. etli bioM resulted in a mutant which took up biotin at a lower rate and accumulated significantly less biotin than the wild type. As in S. meliloti, the R. etli bioMN gene-products resemble the ATPase and permease components, respectively, of an ABC-type transporter. The bioB gene product is in fact similar to members of the BioY family, which has been postulated to function in biotin transport, and we refer to this gene as bioY. An R. etli bioY mutant exhibited lower biotin uptake than the wild-type, providing the first experimental evidence for a role of BioY in biotin transport. We show that the bioMNY operon is transcriptionally repressed by biotin. An analysis of the competitiveness of the wild-type strain versus the bioM mutant showed that the mutant had a diminished capacity to form nodules on bean plants.
由于费氏中华根瘤菌CE3通常依赖于生物素的外部来源且缺乏正统的生物素生物合成基因,我们对该生物体中生物素的摄取进行了分析。通过对苜蓿中华根瘤菌bioM突变体的互补作用,我们分离出了费氏中华根瘤菌的一个染色体区域,该区域编码苜蓿中华根瘤菌bioMNB基因的同源物,其产物与该生物体中生物素的细胞内保留有关。费氏中华根瘤菌bioM的破坏导致一个突变体,其摄取生物素的速率低于野生型,且积累的生物素明显少于野生型。与苜蓿中华根瘤菌一样,费氏中华根瘤菌的bioMN基因产物分别类似于ABC型转运蛋白的ATP酶和通透酶成分。bioB基因产物实际上类似于BioY家族的成员,据推测该家族在生物素转运中起作用,我们将该基因称为bioY。费氏中华根瘤菌bioY突变体的生物素摄取量低于野生型,这为BioY在生物素转运中的作用提供了首个实验证据。我们表明,bioMNY操纵子受到生物素的转录抑制。对野生型菌株与bioM突变体的竞争力分析表明,该突变体在豆类植物上形成根瘤的能力减弱。