Expression of a human 7S K RNA gene in vivo requires a novel pol III upstream element.
The 5'-flanking sequences required for expression of a human 7S K RNA gene have been defined by mutant analysis. A -111 upstream deletion mutant showed full activity when analysed by in vitro transcription with HeLa cell extracts. In contrast, upon transfection into intact cells, this mutant only revealed a basal level activity of approximately 6% as compared to the wild-type promoter up to position -252. The deleted upstream sequence element acts as a transcriptional activator in vivo, in a strictly position and orientation-dependent manner. Two octamer-like binding motifs observed within this upstream sequence were both dispensable for proper function of this RNA polymerase III promoter in vivo. Instead, a detailed analysis of this region identified a CACCC-box element, together with its surrounding base pairs, as the essential upstream element required for expression of this 7S K RNA gene in vivo. Furthermore, this CACCC-box is centered within a footprint obtained with HeLa cell nuclear proteins.
Kleinert
H
H
Department of Biochemistry, Faculty of Chemistry, Ruhr-University, Bochum, FRG.
Bredow
S
S
Benecke
B J
BJ
D001483:Base Sequence; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D005786:Gene Expression Regulation; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D012320:RNA Polymerase III / Q000378:metabolism*; D015183:Restriction Mapping; D012261:Ribonucleoproteins / Q000235:genetics*; D017411:Ribonucleoproteins, Small Nuclear; D014158:Transcription, Genetic; D014162:Transfection
D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear; D012321:DNA-Directed RNA Polymerases; D012320:RNA Polymerase III