Cavicchioli R, Kolesnikow T, Chiang R C, Gunsalus R P
Department of Microbiology and Molecular Genetics, University of California, Los Angeles 90095-1489, USA.
J Bacteriol. 1996 Dec;178(23):6968-74. doi: 10.1128/jb.178.23.6968-6974.1996.
Analysis of the DNA sequence upstream of the narQ gene, which encodes the second nitrate-responsive sensor-transmitter protein in Escherichia coli, revealed an open reading frame (ORF) whose product shows a high degree of similarity to a number of iron-sulfur proteins as well as to the beta subunit of glutamate synthase (gltD) of E. coli. This ORF, located at 53.0 min on the E. coli chromosome, is divergently transcribed and is separated by 206 bp from the narQ gene. Because of the small size of the intergenic region, we reasoned that the genes may be of related function and/or regulated in a similar fashion. An aegA-lacZ gene fusion was constructed and examined in vivo; aegA expression was induced 11-fold by anaerobiosis and repressed 5-fold by nitrate. This control was mediated by the fnr, narX, narQ, and narL gene products. Analysis of an aegA mutant indicated that the aegA gene product is not essential for cell respiration or fermentation or for the utilization of ammonium or the amino acids L-alanine, L-arginine, L-glutamic acid, glycine, and DL-serine as sole nitrogen sources. The ORF was designated aegA to reflect that it is an anaerobically expressed gene. The structural properties of the predicted AegA amino acid sequence and the regulation of aegA are discussed with regard to the possible function of aegA in E. coli.
对narQ基因上游的DNA序列进行分析,该基因编码大肠杆菌中第二种硝酸盐响应传感器-传递蛋白,结果发现一个开放阅读框(ORF),其产物与多种铁硫蛋白以及大肠杆菌谷氨酸合酶(gltD)的β亚基具有高度相似性。这个ORF位于大肠杆菌染色体上53.0分钟处,其转录方向与narQ基因相反,且与narQ基因相隔206 bp。由于基因间区域较小,我们推测这两个基因可能具有相关功能和/或以相似方式受到调控。构建了一个aegA-lacZ基因融合体并在体内进行检测;厌氧条件下aegA表达被诱导11倍,硝酸盐存在时被抑制5倍。这种调控由fnr、narX、narQ和narL基因产物介导。对aegA突变体的分析表明,aegA基因产物对于细胞呼吸或发酵、对于利用铵或作为唯一氮源的L-丙氨酸、L-精氨酸、L-谷氨酸、甘氨酸和DL-丝氨酸等氨基酸并非必需。该ORF被命名为aegA,以反映它是一个厌氧表达的基因。关于aegA在大肠杆菌中的可能功能,讨论了预测的AegA氨基酸序列的结构特性以及aegA的调控情况。