Nasoff M S, Baker H V, Wolf R E
Gene. 1984 Mar;27(3):253-64. doi: 10.1016/0378-1119(84)90070-2.
Expression of gnd of Escherichia coli, which encodes 6-phosphogluconate dehydrogenase, an enzyme of the hexose monophosphate shunt, is subject to growth rate-dependent regulation and is gene dosage-dependent: the level of the enzyme increases in direct proportion to the cellular growth rate at both low and high gene copy numbers. We have determined the nucleotide sequence of gnd and flanking control regions, the 5'-end of in vivo gnd mRNA, and the start codon of the structural gene. Analysis of the sequence indicated that: (i) the gnd promoter is typical of other E. coli promoters and the structural gene is followed by a rho-independent transcription termination signal; (ii) the 56-nucleotide leader of gnd mRNA does not contain a rho-independent transcription termination signal, so growth rate-dependent regulation of 6-phosphogluconate dehydrogenase level is not carried out by an attenuation mechanism analogous to the one that controls expression of the E. coli ampC gene; (iii) the codon composition of the structural gene resembles that of other highly expressed E. coli genes and thus is not responsible for the regulation either; (iv) the structural gene is preceded at an optimal distance by a strong Shine-Dalgarno (SD) sequence, AGGAG ; (v) the leader region of the mRNA contains regions of dyad symmetry that have the potential to sequester the SD sequence and the start codon. This latter feature of the gene suggests that growth rate-dependent regulation may involve regulation of translation initiation frequency.
大肠杆菌中编码6-磷酸葡萄糖酸脱氢酶(磷酸戊糖途径中的一种酶)的gnd基因的表达受生长速率依赖性调控且与基因剂量有关:在低基因拷贝数和高基因拷贝数情况下,该酶的水平均与细胞生长速率成正比增加。我们已确定了gnd及其侧翼控制区的核苷酸序列、体内gnd mRNA的5'端以及结构基因的起始密码子。序列分析表明:(i)gnd启动子是典型的大肠杆菌启动子,结构基因后接一个不依赖ρ因子的转录终止信号;(ii)gnd mRNA的56个核苷酸的前导序列不包含不依赖ρ因子的转录终止信号,因此6-磷酸葡萄糖酸脱氢酶水平的生长速率依赖性调控不是通过类似于控制大肠杆菌ampC基因表达的衰减机制来实现的;(iii)结构基因的密码子组成与其他高表达大肠杆菌基因的相似,因此也不是调控的原因;(iv)结构基因之前在最佳距离处有一个强的Shine-Dalgarno(SD)序列AGGAG;(v)mRNA的前导区包含具有潜在隔离SD序列和起始密码子能力的二元对称区域。该基因的这一特征表明生长速率依赖性调控可能涉及翻译起始频率的调控。