Characterization of the Promoter Regulatory Region of the Human Pyruvate Dehydrogenase .beta. Gene

Madhusudhan, K. T.; Naik, Sharon; Patel, Mulchand S.

doi:10.1021/bi00004a023

Cited by 14 publications

(6 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A 25-fold level of stimulation was observed in K562 cells when the enhancer was located upstream of the heterologous tk promoter and oriented in the same direction as the promoter but only 4-fold in the reverse orientation. Other enhancers have been reported where the extent of functional activity is dependent upon the orientation of the enhancer in the DNA construct (37)(38).…”

Section: Discussionmentioning

confidence: 99%

Identification and Characterization of a Conserved Erythroid-specific Enhancer Located in Intron 8 of the Human 5-Aminolevulinate Synthase 2 Gene

Surinya¹,

Cox²,

May³

1998

Journal of Biological Chemistry

View full text Add to dashboard Cite

Thirty five kilobases of sequence encompassing the human erythroid 5-aminolevulinate synthase (ALAS2) gene have been determined. Analysis revealed a very low GC content, few repetitive elements, and evidence for the insertion of a reverse-transcribed mRNA sequence and a neighboring gene. We have investigated whether introns 1, 3, and 8, which correspond to DNase I-hypersensitivity sites in the structurally related mouse ALAS2 gene, affect expression of the human ALAS2 promoter in transient expression assays. Whereas intron 3 was marginally inhibitory, introns 1 and 8 of the human gene stimulated promoter activity. Intron 8 harbored a strong erythroid-specific enhancer activity which was orientation-dependent. Deletion analysis of this region localized enhancer activity to a fragment of 239 base pairs. Transcription factor binding sites clustered within this region include GATA motifs and CACCC boxes, critical regulatory sequences of many erythroid cell-expressed genes. These sites were also identified in the corresponding intron of both the murine and canine ALAS2 genes. Mutagenesis of these conserved sites in the human intron 8 sequence and transient expression analysis in erythroid cells established the functional importance of one GATA motif and two CACCC boxes. The GATA motif bound GATA-1 in vitro. The two functional CACCC boxes each bound Sp1 or a related protein in vitro, but binding of the erythroid Krü ppel-like factor and the basic Krü ppel-like factor could not be detected. The intron 8 enhancer region was not activated by GATA-1 together with Sp1 in transactivation experiments in COS-1 cells indicating the involvement of a related Sp1 protein or of another unidentified erythroid factor. Overall, these results demonstrate that a GATA-1-binding site and CACCC boxes located within the human ALAS2 intron 8 are critical for the erythroid-specific enhancer activity in transfected erythroid cells, and due to the conserved nature of these binding sites across species, it seems likely that these sites play a functional role in the tissuerestricted expression of the gene in vivo.

show abstract

Section: Discussionmentioning

confidence: 99%

Identification and Characterization of a Conserved Erythroid-specific Enhancer Located in Intron 8 of the Human 5-Aminolevulinate Synthase 2 Gene

Surinya¹,

Cox²,

May³

1998

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…These included one site each for LVa-RS, GRuteroglobin, H4TF-1hist, IE1.2 and two sites for WAP-US6. Some of these sites, such as the H4TF-1, have also been found in the promoter regions of other genes [51,52]. However, it is not known if any of these have functional relevance in the H genes.…”

Section: Discussionmentioning

confidence: 99%

Characterization of the 5′ Flanking Region of the Human Complement Factor H Gene

Williams

Vik

1997

Scand J Immunol

View full text Add to dashboard Cite

Williams SA, Vik DP. Characterization of the 5 0 Flanking Region of the Human Complement Factor H Gene. Scand J Immunol 1997;45:7-15 The promoter region of the human factor H gene was cloned and a 3 kb Eco RI fragment was sequenced. Primer extension and S1 nuclease analysis were used to determine the transcription start site, which was found to be 10-11 nucleotides upstream of the published cDNA sequence. No canonical TATA or CCAAT boxes were found in conjunction with this site. The sequence from the human H promoter region was compared to that from the mouse gene. There was a region of 800 bp that was 62.5% identical between the two sequences. The sequences of the two promoter regions were compared to a database of transcription factor binding sites. Five elements were identified that matched the consensus sequence 100% and were identical in the two promoter sequences. Promoter assays using the luciferase reporter gene demonstrated that this region contained a functional transcription start site and putative enhancer elements. U118-MG astroglioma cells and Hep3b hepatoma cells were incubated with various cytokines to measure effects on their factor H mRNA levels. Interferon-(IFN-), but not interleukin-1 (IL-1), tumour necrosis factor (TNF-) or IL-6, was able to increase the level of H mRNA in both cell lines.

show abstract

“…The only sequence homology in this region to known transcription factor binding sites was to the histone 4 element. Although the H4TF-1 binding site was first identified in the histone H4 gene [36], it has also been found in the human eosinophil peroxidase gene [46] and the human pyruvate dehydrogenase gene [47]. Whether the same nuclear factor is binding to all three sites in these three genes or a different factor for each gene is binding to the same sequence is unknown.…”

Section: Discussionmentioning

confidence: 99%

Regulation of Expression of the Complement Factor H Gene in a Murine Liver Cell Line by Interferon‐γ

Vik

1996

Scand J Immunol

View full text Add to dashboard Cite

Factor H is a regulatory protein of the alternative pathway of complement activation that is synthesized mainly in the liver. The authors used the +/+ Li murine liver cell line as a model for examining its regulation. When +/+ Li cells were incubated with IFN-gamma, the levels of factor H mRNA increased in a dose-dependent manner, achieving a maximal response at a concentration of 50-100 units/ml. The increase in factor H mRNA levels was paralleled by an increase in factor H secretion. The kinetics of induction of factor H mRNA were slow, with the response reaching near maximal levels at 24 h. The increase in factor H mRNA by IFN-gamma was dependent on protein synthesis, as cycloheximide abolished the response. The presence of IFN-gamma was required for the entire incubation period in order to produce a maximal response. The luciferase system was used in an attempt to identify an interferon-responsive element. Luciferase constructs containing from 807 to 236 bp of upstream sequence responded to IFN-gamma with a twofold induction of luciferase activity, whereas a construct containing 83 bp of 5' sequence did not. Thus, IFN-gamma stimulates factor H mRNA transcription through a protein intermediary that interacts with the promoter between positions -83 and -236.

show abstract

Characterization of the Promoter Regulatory Region of the Human Pyruvate Dehydrogenase .beta. Gene

Cited by 14 publications

References 34 publications

Identification and Characterization of a Conserved Erythroid-specific Enhancer Located in Intron 8 of the Human 5-Aminolevulinate Synthase 2 Gene

Identification and Characterization of a Conserved Erythroid-specific Enhancer Located in Intron 8 of the Human 5-Aminolevulinate Synthase 2 Gene

Characterization of the 5′ Flanking Region of the Human Complement Factor H Gene

Regulation of Expression of the Complement Factor H Gene in a Murine Liver Cell Line by Interferon‐γ

Contact Info

Product

Resources

About