Hartmann R K, Ulbrich N, Erdmann V A
Freie Universität Berlin, Institut für Biochemie, FRG.
Nucleic Acids Res. 1987 Oct 12;15(19):7735-47. doi: 10.1093/nar/15.19.7735.
Nuclease S1 mapping analyses were performed in order to detect processing intermediates of pre-23S rRNA from Thermus thermophilus HB8. Two processing sites were identified downstream the start of transcription and several consecutive cleavage sites are associated with the mature 5'-end. In the 3'-flanking region one "primary" site and two cleavages which generate short-living intermediates were detected. A series of successive intermediates in the region of the mature 3'-end implies the existence of--in analogy to Escherichia coli--a 3'-exonucleolytic activity. The data were correlated with potential secondary structures within the pre-23S rRNA, which exhibit various repeated sequence elements. M13 sequencing data support the existence of one secondary structural element associated with the strong "primary" cleavage site in the 3'-flanking region. In T. thermophilus we can exclude the formation of an extended base-paired and precursor-specific stem enclosing the 23S rRNA which is inferred to mediate recognition by RNase III in E. coli.
为了检测嗜热栖热菌HB8的前23S rRNA的加工中间体,进行了核酸酶S1图谱分析。在转录起始位点下游鉴定出两个加工位点,并且几个连续的切割位点与成熟的5'末端相关。在3'侧翼区域检测到一个“主要”位点和两个产生短寿命中间体的切割位点。成熟3'末端区域的一系列连续中间体意味着——类似于大肠杆菌——存在3'外切核酸酶活性。这些数据与前23S rRNA内的潜在二级结构相关,该二级结构呈现出各种重复序列元件。M13测序数据支持在3'侧翼区域存在一个与强“主要”切割位点相关的二级结构元件。在嗜热栖热菌中,我们可以排除形成包围23S rRNA的延伸碱基配对且前体特异性茎的可能性,而在大肠杆菌中,这种茎被推断为介导RNase III的识别。