The housekeeping promoter from the mouse CpG island HTF9 contains multiple protein-binding elements that are functionally redundant.
The mouse CpG-rich island HTF9 harbours the divergent RNA initiation sites shared by two genes that are both expressed in a housekeeping fashion. In this work we have analyzed the architecture of the HTF9 promoter. Gel shift assays were first employed to locate nuclear factor-binding sites within HTF9. Multiple protein-binding sites were identified across a 500 bp-long region, two of which appear to interact with novel factors. Deletion analysis was used to determine the requirements for the different sites in transient expression of a CAT reporter gene. Although multiple elements contributed to the overall promoter strength in each orientation, extensive deletions failed to affect the basal level of transcription from HTF9 in either direction. Thus, only a subset of elements is necessary to activate transcription from HTF9. Functional redundancy may be a general feature of housekeeping CpG-rich promoters.
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Dipartimento di Genetica e Biologia Molecolare, Università La Sapienza, Rome, Italy.
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D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D015139:Blotting, Southern; D015153:Blotting, Western; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002872:Chromosome Deletion; D016172:DNA Fingerprinting; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015226:Dinucleoside Phosphates / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012313:RNA / Q000235:genetics; D014158:Transcription, Genetic
D015500:Chloramphenicol O-Acetyltransferase; D004268:DNA-Binding Proteins; D015226:Dinucleoside Phosphates; D012313:RNA; C015772:CpG dinucleotide