Huang W, Pruzan R, Flint S J
Department of Molecular Biology, Princeton University, NJ 08544.
Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1265-9. doi: 10.1073/pnas.91.4.1265.
We have previously reported that the subgroup C adenovirus E2 early (E2E) RNA polymerase II promoter can specify efficient in vitro transcription by RNA polymerase III. We now show that promoter proximal sequences of the E2E transcription unit are also transcribed by RNA polymerase III in nuclei isolated from adenovirus-infected cells. Small E2E RNA species that possessed the same properties as in vitro synthesized RNA polymerase III E2E transcripts were detected in cytoplasmic RNA populations from infected cells by using blotting, primer extension, and RNase protection assays. The 3' termini of these RNAs were mapped to thymidine-rich sequences typical of RNA polymerase III termination sites. These results demonstrate that a single gene can be transcribed by both RNA polymerase II and RNA polymerase III in vivo.
我们之前报道过,C亚组腺病毒E2早期(E2E)RNA聚合酶II启动子能够在体外通过RNA聚合酶III实现高效转录。我们现在表明,E2E转录单元的启动子近端序列在从腺病毒感染细胞中分离出的细胞核内也由RNA聚合酶III转录。通过印迹法、引物延伸法和核糖核酸酶保护试验,在来自感染细胞的细胞质RNA群体中检测到了具有与体外合成的RNA聚合酶III E2E转录本相同特性的小E2E RNA种类。这些RNA的3'末端被定位到富含胸腺嘧啶的序列,这是RNA聚合酶III终止位点的典型特征。这些结果表明,单个基因在体内可由RNA聚合酶II和RNA聚合酶III转录。