Kessler Alexandra, Sezonov Guennadi, Guijarro J Iñaki, Desnoues Nicole, Rose Thierry, Delepierre Muriel, Bell Stephen D, Prangishvili David
Unité de Biologie Moléculaire du Gène chez les Extrêmophiles, Institut Pasteur, 25-28 rue du Dr Roux, 75724 Paris Cedex 15, France.
Nucleic Acids Res. 2006;34(17):4837-45. doi: 10.1093/nar/gkl502. Epub 2006 Sep 14.
While studying gene expression of the rudivirus SIRV1 in cells of its host, the hyperthermophilic crenarchaeon Sulfolobus, a novel archaeal transcriptional regulator was isolated. The 14 kDa protein, termed Sulfolobus transcription activator 1, Sta1, is encoded on the host chromosome. Its activating effect on transcription initiation from viral promoters was demonstrated in in vitro transcription experiments using a reconstituted host system containing the RNA polymerase, TATA-binding protein (TBP) and transcription factor B (TFB). Most pronounced activation was observed at low concentrations of either of the two transcription factors, TBP or TFB. Sta1 was able to bind viral promoters independently of any component of the host pre-initiation complex. Two binding sites were revealed by footprinting, one located in the core promoter region and the second approximately 30 bp upstream of it. Comparative modeling, NMR and circular dichroism of Sta1 indicated that the protein contained a winged helix-turn-helix motif, most probably involved in DNA binding. This strategy of the archaeal virus to co-opt a host cell regulator to promote transcription of its genes resembles eukaryal virus-host relationships.
在研究嗜热泉古菌硫化叶菌(Sulfolobus)细胞中rudivirus病毒SIRV1的基因表达时,分离出了一种新型古菌转录调节因子。这种14 kDa的蛋白质被称为硫化叶菌转录激活因子1(Sta1),由宿主染色体编码。在使用包含RNA聚合酶、TATA结合蛋白(TBP)和转录因子B(TFB)的重组宿主系统进行的体外转录实验中,证明了它对病毒启动子转录起始的激活作用。在两种转录因子TBP或TFB浓度较低时,观察到了最明显的激活作用。Sta1能够独立于宿主预起始复合物的任何成分与病毒启动子结合。足迹法揭示了两个结合位点,一个位于核心启动子区域,另一个在其上游约30 bp处。Sta1的比较建模、核磁共振和圆二色性表明,该蛋白质含有一个翼状螺旋-转角-螺旋基序,很可能参与DNA结合。古菌病毒利用宿主细胞调节因子促进其基因转录的这种策略类似于真核病毒与宿主的关系。