Matsutani Sachiko
Division of Microbiology, National Institute of Health Sciences, Tokyo, 158-8501, Japan.
Genetica. 2007 Oct;131(2):127-34. doi: 10.1007/s10709-006-9122-5. Epub 2006 Dec 7.
The bacterial repetitive sequence IS1, is a translocatable DNA segment. The internal region of IS1 acts as a cis-element to stimulate RNA synthesis from the upstream promoter. The product of the bacterial artA gene works with this cis-element to stimulate transcription. Eukaryotic genes for small RNAs and short interspersed repetitive elements (SINEs) have internal promoters, transcribed by RNA polymerase III (RNAP III). RNAP III requires the multisubunit protein factor TFIIIC in transcription initiation. TFIIIC contains the B-block binding subunit which recognizes the internal promoter. Here, I report that the eukaryotic RNAP III promoter-like sequence was found in the cis-element of bacterial IS1. Mutations in the cis-element which affect transcription were present in the RNAP III promoter-like sequence. The RNAP III promoter sequence of Alu, which is a human SINE, was cloned into Escherichia coli, and was shown to stimulate bacterial transcription like the cis-element of IS1. Furthermore, the primary structures of ArtA protein and B-block binding subunits were compared. The amino acid sequence of ArtA appeared to be similar to the N- and C-terminal regions conserved in many B-block binding subunits. Prokaryotes and eukaryotes have been thought to have inherent transcription machineries. The results shown here, however, suggest a new aspect of the evolution of the RNAP III transcription machinery.
细菌重复序列IS1是一个可转位的DNA片段。IS1的内部区域作为顺式元件,刺激上游启动子的RNA合成。细菌artA基因的产物与该顺式元件共同作用以刺激转录。小RNA和短散在重复元件(SINEs)的真核基因具有内部启动子,由RNA聚合酶III(RNAP III)转录。RNAP III在转录起始时需要多亚基蛋白因子TFIIIC。TFIIIC包含识别内部启动子的B块结合亚基。在此,我报告在细菌IS1的顺式元件中发现了真核RNAP III启动子样序列。影响转录的顺式元件中的突变存在于RNAP III启动子样序列中。人类SINE Alu的RNAP III启动子序列被克隆到大肠杆菌中,并显示出像IS1的顺式元件一样刺激细菌转录。此外,比较了ArtA蛋白和B块结合亚基的一级结构。ArtA的氨基酸序列似乎与许多B块结合亚基中保守的N端和C端区域相似。原核生物和真核生物一直被认为具有固有的转录机制。然而,此处所示结果提示了RNAP III转录机制进化的一个新方面。