In vivo activity of the most proximal promoter of the human aldolase A gene and analysis of transcriptional control elements.
The genomic region upstream from exon F (exon IV) of the human aldolase A gene has been studied for its ability to direct the transcription of a reporter gene in vivo. Transfection experiments in human hepatoma cells (Hep 3B) followed by CAT assay, and S1 mapping analysis, demonstrated that: (i) this region is able to drive CAT gene transcription; (ii) all the transcriptional control elements of this promoter are downstream from nucleotide -384 of the longer ubiquitous RNA start site and the sequences between -384 and -262 play a crucial role in transcriptional efficiency; (iii) initiation starting points for two mRNAs exist 61 bp apart. Gel retardation and footprinting assays demonstrated the presence of DNA-protein complexes mainly in the region between -384 and -262 and such ubiquitous binding factors as Sp1 and AP-1.
Izzo
P
P
Department of Biochemistry and Medical Biotechnology, Faculty of Medicine II, University of Naples, Italy.
Costanzo
P
P
Lupo
A
A
Rippa
E
E
Salvatore
F
F
D001483:Base Sequence; D006528:Carcinoma, Hepatocellular; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002872:Chromosome Deletion; D003850:Deoxyribonuclease I; D005091:Exons; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics*; D005796:Genes; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008113:Liver Neoplasms; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014162:Transfection
D009687:Nuclear Proteins; D015500:Chloramphenicol O-Acetyltransferase; D003850:Deoxyribonuclease I; D005634:Fructose-Bisphosphate Aldolase