Kjems J, Leffers H, Garrett R A, Wich G, Leinfelder W, Böck A
Nucleic Acids Res. 1987 Jun 25;15(12):4821-35. doi: 10.1093/nar/15.12.4821.
The single ribosomal RNA (rRNA) operon from the extreme thermophile and archaebacterium Thermoproteus tenax was sequenced. Sites of transcriptional initiation and termination were established and the processing sites on the primary transcript were mapped with nuclease S1. The operon contained genes coding for 16S and 23S RNAs but lacked those coding for tRNA and 5S RNA. Transcription initiates 175 bp upstream from the start of the 16S RNA gene (Wich et al., EMBO J. 6, 523-528, 1987) and terminates 49 bp downstream from the 23S RNA gene within a long pyrimidine sequence. An open reading frame downstream from the rRNA operon is transcribed. The sequences bordering both 16S and 23S RNA genes can form putative processing stems in the primary transcript that involve the whole of the 16S-23S RNA spacer. The stems contain irregular features that constitute processing signals and are conserved in other archaebacteria. The 16S RNA stem is cut prior to that of the 23S RNA and RNA maturation follows. An unusual 14 bp helix can form between the extremities of the transcript such that the whole transcript is highly structured and a fork-like structure is formed together with the processing stems. The 23S RNA sequence was aligned with other available 23S-like RNA sequences (Leffers et al., J. Mol. Biol. 195, in press): a putative secondary structure exhibiting archaebacterial-specific features was deduced using comparative sequence analyses. A rooted phylogenetic tree was also derived for the archaebacteria that confirms their division into three major subgroups.
对嗜热栖热袍菌(一种极端嗜热古细菌)的单个核糖体RNA(rRNA)操纵子进行了测序。确定了转录起始和终止位点,并用核酸酶S1定位了初级转录本上的加工位点。该操纵子包含编码16S和23S RNA的基因,但缺少编码tRNA和5S RNA的基因。转录起始于16S RNA基因起始位点上游175 bp处(Wich等人,《欧洲分子生物学组织杂志》6,523 - 528,1987),并在23S RNA基因下游49 bp处的一个长嘧啶序列内终止。rRNA操纵子下游的一个开放阅读框被转录。与16S和23S RNA基因相邻的序列可在初级转录本中形成推测的加工茎,该茎涉及整个16S - 23S RNA间隔区。这些茎包含构成加工信号的不规则特征,并且在其他古细菌中保守。16S RNA茎在23S RNA茎之前被切割,随后进行RNA成熟。转录本末端之间可形成一个不寻常的14 bp螺旋,使得整个转录本具有高度结构化,并与加工茎一起形成叉状结构。将23S RNA序列与其他可用的23S样RNA序列进行比对(Leffers等人,《分子生物学杂志》195,即将发表):通过比较序列分析推导了一个具有古细菌特异性特征的推测二级结构。还为古细菌构建了一个有根系统发育树,证实它们分为三个主要亚组。