Hausner W, Lange U, Musfeldt M
Institut für Allgemeine Mikrobiologie, University of Kiel, Am Botanischen Garten 1-9, D-24118 Kiel, Federal Republic of Germany.
J Biol Chem. 2000 Apr 28;275(17):12393-9. doi: 10.1074/jbc.275.17.12393.
We have analyzed the function of an archaeal protein (now called transcription factor S (TFS)) that shows sequence similarity to eukaryotic transcription factor IIS (TFIIS) as well as to small subunits of eukaryotic RNA polymerases I (A12.6), II (B12.2), and III (C11). Western blot analysis with antibodies against recombinant TFS demonstrated that this protein is not a subunit of the RNA polymerase. In vitro transcription experiments with paused elongation complexes at position +25 showed that TFS is able to induce cleavage activity in the archaeal RNA polymerase in a similar manner to TFIIS. In the presence of TFS, the cleavage activity of the RNA polymerase truncates the RNA back to position +15 by releasing mainly dinucleotides from the 3'-end of the nascent RNA. Furthermore, TFS reduces the amount of non-chaseable elongation complexes at position +25 as well as position +45. These findings clearly demonstrate that this protein has a similar function to eukaryotic TFIIS.
我们分析了一种古细菌蛋白(现称为转录因子S(TFS))的功能,该蛋白与真核转录因子IIS(TFIIS)以及真核RNA聚合酶I(A12.6)、II(B12.2)和III(C11)的小亚基具有序列相似性。用抗重组TFS的抗体进行的蛋白质印迹分析表明,该蛋白不是RNA聚合酶的亚基。在+25位具有暂停延伸复合物的体外转录实验表明,TFS能够以与TFIIS类似的方式诱导古细菌RNA聚合酶的切割活性。在TFS存在的情况下,RNA聚合酶的切割活性通过从新生RNA的3'末端主要释放二核苷酸将RNA截短至+15位。此外,TFS减少了+25位以及+45位不可追踪延伸复合物的数量。这些发现清楚地表明,该蛋白具有与真核TFIIS相似的功能。