Archaeal Genetics Laboratory, Department of Molecular Biology, University of Copenhagen, Copenhagen Biocenter Ole Maaløes Vej 5, 2200, Copenhagen N, Denmark.
Extremophiles. 2010 Sep;14(5):453-63. doi: 10.1007/s00792-010-0325-4. Epub 2010 Aug 25.
The pSSVx from Sulfolobus islandicus, strain REY15/4, is a hybrid between a plasmid and a fusellovirus. A systematic study previously performed revealed the presence of nine major transcripts, the expression of which was differentially and temporally regulated over the growth cycle of S. islandicus. In this study, two new transcripts were identified. Then, 3' termini of all the RNAs were mapped using adaptor RT-PCR and RNase protection assays, and termination/arrest positions were identified for each transcript. The majority of the identified ending positions were located in the close vicinity of a T-rich sequence and this was consistent with termination signals identifiable for most of archaeal genes. Furthermore, termination also occurred at locations where a T-track sequence was absent but a stem-loop structure could be formed. We propose that an alternative mechanism based on secondary RNA structures and counter-transcripts might be responsible for the transcription termination at these T-track-minus loci in the closely spaced pSSVx genes.
来自嗜酸热硫化叶菌(Sulfolobus islandicus)菌株 REY15/4 的 pSSVx 是质粒和丝状病毒之间的杂种。之前进行的一项系统研究表明,存在 9 种主要转录本,它们的表达在嗜酸热硫化叶菌的生长周期中表现出差异和时间调节。在这项研究中,鉴定出了两个新的转录本。然后,使用接头 RT-PCR 和 RNase 保护分析对所有 RNA 的 3' 末端进行了映射,并确定了每个转录本的终止/逮捕位置。大多数鉴定的终止位置位于富含 T 的序列附近,这与大多数古细菌基因的终止信号一致。此外,终止也发生在不存在 T 轨道序列但可以形成茎环结构的位置。我们提出,基于二级 RNA 结构和反式转录本的替代机制可能负责在这些紧密间隔的 pSSVx 基因中 T 轨道缺失位点的转录终止。