Myslinski E, Krol A, Carbon P
Institut de Biologie Moléculaire et Cellulaire du CNRS, Strasbourg, France.
Nucleic Acids Res. 1992 Jan 25;20(2):203-9. doi: 10.1093/nar/20.2.203.
The X. laevis tRNA((Ser)Sec) gene is different from the other tRNA genes in that its promoter contains two external elements, a PSE and a TATA box functionally equivalent to those of the U6 snRNA gene. Of the two internal promoters governing classical tRNA gene transcription, only subsists the internal B box. In this report, we show that the tRNA((Ser)Sec) contains in addition an activator element (AE) which we have mapped by extensive mutagenesis. Activation is only dependent on a 15 bp fragment residing between -209 and -195 and containing an SPH motif. In vitro, this element forms a complex with a nuclear protein which is different from the TEF-1 transcriptional activator that binds the SV40 Sph motifs. This AE is versatile since it shows capacity of activating a variety of genes in vivo, including U1 and U6 snRNAs and HSV thymidine kinase. Unexpectedly for an snRNA-related gene, the tRNA((Ser)Sec) is deprived of octamer or octamer-like motifs. The X.laevis tRNA((Ser)Sec) gene represents the first example of a Pol III snRNA-type gene whose activation of transcription is completely octamer-independent.
非洲爪蟾tRNA((Ser)Sec)基因与其他tRNA基因不同,其启动子包含两个外部元件,一个近端序列元件(PSE)和一个功能上等同于U6小核RNA(snRNA)基因启动子元件的TATA框。在控制经典tRNA基因转录的两个内部启动子中,仅保留了内部B框。在本报告中,我们表明tRNA((Ser)Sec)还包含一个激活元件(AE),我们通过广泛的诱变对其进行了定位。激活仅依赖于位于-209至-195之间且包含SPH基序的15个碱基对片段。在体外,该元件与一种核蛋白形成复合物,该核蛋白不同于结合SV40 Sph基序的TEF-1转录激活因子。这个AE具有通用性,因为它在体内显示出激活多种基因的能力,包括U1和U6 snRNA以及单纯疱疹病毒胸苷激酶。对于一个与snRNA相关的基因而言出乎意料的是,tRNA((Ser)Sec)缺乏八聚体或类八聚体基序。非洲爪蟾tRNA((Ser)Sec)基因代表了第一个转录激活完全不依赖八聚体的RNA聚合酶III snRNA型基因的例子。