Sacerdot C, Dessen P, Hershey J W, Plumbridge J A, Grunberg-Manago M
Proc Natl Acad Sci U S A. 1984 Dec;81(24):7787-91. doi: 10.1073/pnas.81.24.7787.
The gene for protein synthesis initiation factor IF2 in Escherichia coli, infB, is located downstream from nusA on the same operon. We sequenced about 3 kilobases of DNA beginning within nusA and including the entire infB structural gene plus another 392 bases downstream. This region contains no obvious strong promoter signals, but a possible transcriptional termination or pausing site occurs downstream from infB. The putative initiator codon for IF2 alpha (97,300 daltons) is AUG; that for IF2 beta (79,700 daltons) is GUG, located 471 bases downstream in the same reading frame. The codon usage for IF2 is typical of other highly expressed proteins in E. coli and suggests that IF2 mRNA is efficiently translated. IF2 alpha contains two adjacent regions (residues 104-155 and 167-214) that are rich in alanine and charged amino acids and that show striking periodicities in their sequences. These regions may alternate between flexible and helical conformations, thereby drawing together the NH2-terminal and COOH-terminal globular domains of the factor as IF2 interacts with ribosomes or tRNA. Certain regions of the DNA and protein sequences of IF2 share strong homologies with elongation factor EF-Tu and lesser homology with EF-G. In particular, a region of EF-Tu implicated in GTP binding contains sequences and secondary structure that are conserved in IF2. The homologies indicate that the genes for IF2 and the elongation factors are derived at least in part from a common ancestor.
大肠杆菌中蛋白质合成起始因子IF2的基因infB位于同一操纵子上nusA的下游。我们对大约3千碱基的DNA进行了测序,起始于nusA内部,包括整个infB结构基因以及下游另外392个碱基。该区域没有明显的强启动子信号,但在infB下游存在一个可能的转录终止或暂停位点。IF2α(97,300道尔顿)的推定起始密码子是AUG;IF2β(79,700道尔顿)的推定起始密码子是GUG,位于同一阅读框下游471个碱基处。IF2的密码子使用情况与大肠杆菌中其他高表达蛋白的情况类似,这表明IF2 mRNA能被有效翻译。IF2α包含两个相邻区域(第104 - 155位残基和第167 - 214位残基),这些区域富含丙氨酸和带电荷的氨基酸,并且其序列呈现出显著的周期性。当IF2与核糖体或tRNA相互作用时,这些区域可能在柔性构象和螺旋构象之间交替,从而将该因子的NH2末端和COOH末端球状结构域聚集在一起。IF2的DNA和蛋白质序列的某些区域与延伸因子EF - Tu具有很强的同源性,与EF - G的同源性较弱。特别是,EF - Tu中与GTP结合相关的一个区域包含在IF2中保守的序列和二级结构。这些同源性表明,IF2基因和延伸因子基因至少部分源自共同的祖先。