title: Brain-specific enhancement of the mouse neurofilament heavy gene promoter in vitro.
abstract: We have investigated the DNA elements responsible for transcription from the proximal portion of the mouse neurofilament heavy gene (NF-H) promoter by in vitro transcription using extracts from expressing (brain) and non-expressing (liver) tissues. We have found that constructs containing 5' region from -1314 to -115 exhibit a 3-5-fold higher level of NF-H promoter activity, relative to the adenovirus major late promoter (pML), in brain versus liver extracts. Deletion to -85 lowers the level of brain transcription by 2-fold, while deletion from -65 through -31 reduces transcription by 5-fold to a relatively strong (10% of pML) basal level. Basal level expression is observed in all deletions transcribed with liver extract. Deletion to -24 (TATA-less) abolishes promoter activity with both extracts. Deletion of the -115 to -65 region from a larger construct reduces transcription in brain extracts to basal levels, suggesting that this region contains the elements necessary for the brain-specific enhancement of promoter function. Mutation of a palindromic sequence within this region abolishes brain-specific enhanced promoter activity. This loss of enhanced transcriptional activity is correlated with the loss of a shifted band in gel shift assays. Our studies suggest that the sequence (-106)GGGGAGGAGG-(15 bp)-CCTCCTCCCC(-72) (where bp = base pairs) is important in brain-specific enhancement of transcription from the mouse NF-H promoter.
authors.affiliation: Division of Neuropathology, University of Pennsylvania Medical School, Philadelphia 19104.
authors.forename: M L
authors.initials: ML
authors.lastname: Schwartz
authors.forename: C
authors.initials: C
authors.lastname: Katagi
authors.forename: J
authors.initials: J
authors.lastname: Bruce
authors.forename: W W
authors.initials: WW
authors.lastname: Schlaepfer
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D004247:DNA; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D016900:Neurofilament Proteins / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D051381:Rats; D014158:Transcription, Genetic
chemical_list: D016900:Neurofilament Proteins; D004247:DNA