Gavin B J, Ward D C
Department of Human Genetics, Yale University School of Medicine, New Haven, Connecticut 06510-8005.
J Virol. 1990 May;64(5):2057-63. doi: 10.1128/JVI.64.5.2057-2063.1990.
5' deletion mutants of the minute virus of mice P38 promoter were constructed and analyzed for transcriptional activity in vitro and in vivo. In uninfected mouse A9 cell extracts, 107 base pairs upstream of the RNA start sites are required for optimal activity. Within this region, the only readily recognizable cis-acting control elements are a GC box and a TATA box. However, in infected cell extracts, deletion of a sequence between -167 and -121, which shares homology with the 30-base-pair trans-activation region (TAR) of H-1 virus (S. L. Rhode and S. M. Richard, J. Virol. 61:2807-2815, 1987), results in a three- to fourfold decrease in transcriptional activity. Interestingly, in vivo transfection experiments demonstrate a three- to eightfold increase in transcription relative to the wild-type promoter when the TAR element homology region is deleted and reveal a functional role for a CCAAT motif which lies immediately downstream of the TAR element. These results indicate both positive and negative regulation of the P38 promoter.
构建了小鼠微小病毒P38启动子的5'缺失突变体,并对其在体外和体内的转录活性进行了分析。在未感染的小鼠A9细胞提取物中,RNA起始位点上游107个碱基对是实现最佳活性所必需的。在该区域内,唯一易于识别的顺式作用控制元件是一个GC盒和一个TATA盒。然而,在感染的细胞提取物中,删除与H-1病毒30个碱基对的反式激活区域(TAR)具有同源性的-167至-121之间的序列(S. L. Rhode和S. M. Richard,《病毒学杂志》61:2807-2815,1987),会导致转录活性降低三到四倍。有趣的是,体内转染实验表明,当删除TAR元件同源区域时,相对于野生型启动子,转录增加三到八倍,并揭示了紧邻TAR元件下游的一个CCAAT基序的功能作用。这些结果表明P38启动子存在正调控和负调控。