Analysis of clustered point mutations in the human ribosomal RNA gene promoter by transient expression in vivo.
We have mapped the cis regulatory elements required in vivo for initiation at the human rRNA promoter by RNA polymerase I. Transient expression in COS-7 cells was used to evaluate the transcription phenotype of clustered base substitution mutations in the human rRNA promoter. The promoter consists of two major elements: a large upstream region, composed of several domains, that lies between nucleotides -234 and -107 relative to the transcription initiation site and affects transcription up to 100-fold and a core element that lies between nucleotides -45 and +20 and affects transcription up to 1000-fold. The upstream region is able to retain partial function when positioned within 100-160 nucleotides of the transcription initiation site, but it cannot stimulate transcription from distances of greater than or equal to 600 nucleotides. In addition, we demonstrate, using mouse-human hybrid rRNA promoters, that the sequences responsible for human species-specific transcription in vivo appear to reside in both the core and upstream elements, and sequences from the mouse rRNA promoter cannot be substituted for them.
Howard Hughes Medical Institute, University of California, Berkeley 94720.
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MH
Jones
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Learned
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Tjian
D000818:Animals; D001483:Base Sequence; D004275:DNA, Ribosomal / Q000235:genetics*; D006801:Humans; D006822:Hybrid Cells / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D012335:RNA, Ribosomal / Q000235:genetics*; D014158:Transcription, Genetic
D004275:DNA, Ribosomal; D012335:RNA, Ribosomal