Hain J, Reiter W D, Hüdepohl U, Zillig W
Max-Planck-Institut für Biochemie, Martinsried, Germany.
Nucleic Acids Res. 1992 Oct 25;20(20):5423-8. doi: 10.1093/nar/20.20.5423.
The sequence requirements for specific and efficient transcription from the 16S/23S rRNA promoter of Sulfolobus shibatae were analysed by point mutations and by cassette mutations using an in vitro transcription system. The examination of the box A-containing distal promoter element (DPE) showed the great importance of the TA sequence in the center of box A for transcription efficiency and the influence of the sequence upstream of box A on determining the distance between the DPE and the start site. In most positions of box A, replacement of the wild type bases by adenines or thymines are less detrimental than replacements by cytosines or guanines. The effectiveness of the proximal promoter element (PPE) was not merely determined by its high A + T content but appeared to be directly related to its nucleotide sequence. At the start site a pyrimidine/purine (py/pu) sequence was necessary for unambiguous initiation as shown by analysis of mutants where the wild type start base was replaced. The sequence of box A optimal for promoter function in vitro is identical to the consensus of 84 mapped archaeal promoter sequences.
利用体外转录系统，通过点突变和盒式突变分析了柴田硫化叶菌16S/23S rRNA启动子进行特异性高效转录的序列要求。对含A框的远端启动子元件（DPE）的研究表明，A框中心的TA序列对转录效率至关重要，且A框上游序列对确定DPE与起始位点之间的距离有影响。在A框的大多数位置，用腺嘌呤或胸腺嘧啶取代野生型碱基的危害小于用胞嘧啶或鸟嘌呤取代。近端启动子元件（PPE）的有效性不仅取决于其高A+T含量，似乎还与其核苷酸序列直接相关。如对野生型起始碱基被取代的突变体分析所示，在起始位点，嘧啶/嘌呤（py/pu）序列对于明确起始是必需的。体外启动子功能的最佳A框序列与84个已定位古菌启动子序列的共有序列相同。
