In vivo activity of the most proximal promoter of the human aldolase A ene and analysis of transcriptional control elements

Abstract

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 S, 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 Spl and AP-1.

The Krüppel-like zinc-finger protein ZNF224 represses aldolase A gene transcription by interacting with the KAP-1 co-repressor protein

“…The availability of soluble ZNF224 molecules and accessibility of its cognate motif on the AldA-NRE in the distal promoter of the aldolase A gene are prerequisites for full ZNF224-mediated transcription repression. Coordinated regulation of aldolase A gene transcription involves multiple cis-elements located in the promoter regions upstream from leader exons L1, M, and F that interact with several nuclear proteins (Izzo et al, 1989;Concordet et al, 1991;Costanzo et al, 1993;Salminen et al, 1994;Lupo et al, 1995Lupo et al, , 1997. A negative regulatory element could determine the timing and distribution of transcription during development.…”

“…Regulation of the aldolase A gene involves multiple ciselements located within the promoter regions upstream from leader exons L1, M and F that interact with several nuclear proteins (Izzo et al, 1989;Costanzo et al, 1993;Concordet et al, 1991;Lupo et al, 1995). The differential expression of various aldolase A mRNAs (L, M and F types) is controlled by interaction between DNA and proteins according to cell and stage specificity (Salminen et al, 1994).…”

Negative Regulation of the Mouse Aldolase A Gene

“…Transcription of multiple mRNA species (L, M, and F types), containing the same coding region but different untranslated 5Ј-ends, is driven by three autonomous promoters and splicing of alternative leader exons (L, M, and F) (1-3). In man, two of these mRNA species (L and F types) are ubiquitously expressed, whereas the third one (M type) is muscle-specific (4 -6).We have demonstrated that the human genomic regions upstream from exons F and L are able to drive autonomous transcription (7,8). We found that the basal transcriptional activity of pF is regulated by several binding sites for the ubiquitous factor Sp1 (7).…”

“…Regulation of the aldolase A gene involves multiple cis-elements located within the promoter regions upstream from leader exons L1, M, and F that interact with several nuclear proteins (7)(8)(9)(10)(11). The differential expression of various aldolase A mRNAs (L, M, and F types) is controlled by an interplay of interactions between DNA and proteins according to cell and stage specificity (20).…”

“…We found that the basal transcriptional activity of pF is regulated by several binding sites for the ubiquitous factor Sp1 (7). Binding of nuclear trans-acting factors AP-1 and NF-1 in the Ϫ384/Ϫ262 region increases pF transcriptional activity (7,9).…”