Sakamoto T, Inoue-Sakamoto K, Bryant D A
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
J Bacteriol. 1999 Dec;181(23):7363-72. doi: 10.1128/JB.181.23.7363-7372.1999.
The nrtP and narB genes, encoding nitrate/nitrite permease and nitrate reductase, respectively, were isolated from the marine cyanobacterium Synechococcus sp. strain PCC 7002 and characterized. NrtP is a member of the major facilitator superfamily and is unrelated to the ATP-binding cassette-type nitrate transporters that previously have been described for freshwater strains of cyanobacteria. However, NrtP is similar to the NRT2-type nitrate transporters found in diverse organisms. An nrtP mutant strain consumes nitrate at a 4.5-fold-lower rate than the wild type, and this mutant grew exponentially on a medium containing 12 mM nitrate at a rate approximately 2-fold lower than that of the wild type. The nrtP mutant cells could not consume nitrite as rapidly as the wild type at pH 10, suggesting that NrtP also functions in nitrite uptake. A narB mutant was unable to grow on a medium containing nitrate as a nitrogen source, although this mutant could grow on media containing urea or nitrite with rates similar to those of the wild type. Exogenously added nitrite enhanced the in vivo activity of nitrite reductase in the narB mutant; this suggests that nitrite acts as a positive effector of nitrite reductase. Transcripts of the nrtP and narB genes were detected in cells grown on nitrate but were not detected in cells grown on urea or ammonia. Transcription of the nrtP and narB genes is probably controlled by the NtcA transcription factor for global nitrogen control. The discovery of a nitrate/nitrite permease in Synechococcus sp. strain PCC 7002 suggests that significant differences in nutrient transporters may occur in marine and freshwater cyanobacteria.
分别编码硝酸盐/亚硝酸盐通透酶和硝酸盐还原酶的nrtP和narB基因,是从海洋蓝藻聚球藻属PCC 7002菌株中分离出来并进行表征的。NrtP是主要转运蛋白超家族的成员,与先前已描述的淡水蓝藻菌株中的ATP结合盒式硝酸盐转运蛋白无关。然而,NrtP与在多种生物体中发现的NRT2型硝酸盐转运蛋白相似。一个nrtP突变株消耗硝酸盐的速率比野生型低4.5倍,并且该突变株在含有12 mM硝酸盐的培养基上指数生长,其速率比野生型低约2倍。在pH 10时,nrtP突变细胞不能像野生型那样快速消耗亚硝酸盐,这表明NrtP在亚硝酸盐摄取中也起作用。一个narB突变株不能在以硝酸盐作为氮源的培养基上生长,尽管该突变株可以在含有尿素或亚硝酸盐的培养基上以与野生型相似的速率生长。外源添加的亚硝酸盐增强了narB突变株中亚硝酸盐还原酶的体内活性;这表明亚硝酸盐作为亚硝酸盐还原酶的正效应物。在以硝酸盐生长的细胞中检测到nrtP和narB基因的转录本,但在以尿素或氨生长的细胞中未检测到。nrtP和narB基因的转录可能受用于全局氮控制的NtcA转录因子调控。在聚球藻属PCC 7002菌株中发现硝酸盐/亚硝酸盐通透酶表明,海洋和淡水蓝藻在营养物质转运蛋白方面可能存在显著差异。