title: An element downstream of the cap site is required for transcription of the gene encoding mouse ribosomal protein L32.
abstract: To identify the elements that regulate transcription of the mouse gene encoding ribosomal protein L32 (rpL32), we transfected monkey kidney (COS or CV-1) cells with mutants bearing progressive 5' deletions or an internal deletion in exon I and measured their transient expression by S1 nuclease protection analysis. When the mutant genes were tested in the vector pi SVHSplac, which contains a short segment of the oriregion of simian virus 40, maximum expression was observed with as little as 36 base pairs of 5' flanking sequence, and the mutant bearing the exon I deletion was expressed very efficiently. However, when the genes were tested in a simple prokaryotic (pUC) vector, the expression was increased 3- to 4-fold by sequences between -36 and -159, and the exon I segment was absolutely required for expression. Gel mobility-shift and methylation interference analyses revealed that a nuclear factor specifically binds to a GGCTGCCATC sequence within this exon I segment. These results, taken together with other recent findings, indicate that the elements involved in transcriptional regulation of the rpL32 gene are distributed over a 200-base-pair region that spans the cap site. The contributions of some of these elements are apparently masked in the presence of simian virus 40 ori-region elements.
authors.affiliation: Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA 19111.
authors.forename: R
authors.initials: R
authors.lastname: Moura-Neto
authors.forename: K P
authors.initials: KP
authors.lastname: Dudov
authors.forename: R P
authors.initials: RP
authors.lastname: Perry
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002872:Chromosome Deletion; D005796:Genes; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012315:RNA Caps / Q000235:genetics*; D015183:Restriction Mapping; D012269:Ribosomal Proteins / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D012315:RNA Caps; D012269:Ribosomal Proteins; C041806:ribosomal protein L32