Cited by 74Hemoglobinopathies and Related DisordersGenomics and Chromatin DynamicsPorphyrin Metabolism and Disorders

Transcriptional Regulation of the Human Erythroid 5-Aminolevulinate Synthase Gene

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Surinya, Katharina H., Cox, Timothy C., May, Brian K.·1997·Journal of Biological Chemistry

Abstract

We have characterized the 5'-flanking region of the human erythroid-specific 5-amino levulinate synthase (ALAS) gene (the ALAS2 gene) and shown that the first 300 base pairs of promoter sequence gives maximal expression in erythroid cells. Transcription factor binding sites clustered within this promoter sequence include GATA motifs and CACCC boxes, critical regulatory sequences of many erythroid cell-expressed genes. GATA sites at -126/-121 (on the noncoding strand) and -102/-97 were each recognized by GATA-1 protein in vitro using erythroid cell nuclear extracts. Promoter mutagenesis and transient expression assays in erythroid cells established that both GATA-1 binding sites were functional and exogenously expressed GATA-1 increased promoter activity through these sites in transactivation experiments. A noncanonical TATA sequence at the expected TATA box location (-30/-23) bound GATA-1- or TATA-binding protein (TBP) in vitro. Conversion of this sequence to a canonical TATA box reduced expression in erythroid cells, suggesting a specific role for GATA-1 at this site. However, expression was also markedly reduced when the -30/-23 sequence was converted to a consensus GATA-1 sequence (that did not bind TBP in vitro), suggesting that a functional interaction of both factors with this sequence is important. A sequence comprising two overlapping CACCC boxes at -59/-48 (on the noncoding strand) was demonstrated by mutagenesis to be functionally important. This CACCC sequence bound Sp1, erythroid Krüppel-like factor, and basic Krüppel-like factor in vitro, while in transactivation experiments erythroid Krüppel-like factor activated ALAS2 promoter expression through this sequence. A sequence at -49/-39 with a 9/11 match to the consensus for the erythroid specific factor NF-E2 was not functional. Promoter constructs with 5'-flanking sequence from 293 base pairs to 10.3 kilobase pairs expressed efficiently in COS-1 cells as well as in erythroid cells, indicating that an enhancer sequence located elsewhere or native chromatin structure may be required for the tissue-restricted expression of the gene in vivo.

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Crossref · 10.1074/jbc.272.42.26585

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  • There is evidence that heme not only regulates ALAS1 mitochondrial import but also attenuates its transcription.

    “Moreover, there is evidence that heme not only regulates ALAS1 mitochondrial import but also attenuates its transcription [36,37].”

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Authors
Surinya, K. H., Cox, T. C., & May, B. K.
Affiliations
The University of Adelaide (AU)
Corresponding author
May, Brian K.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1997-10
Volume
272
Issue
42
Pages
26585-26594
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2083397383
Cited by
74
References
72
Citations by year
2026: 1 · 2025: 1 · 2024: 3 · 2023: 1 · 2022: 2 · 2020: 2 · 2018: 1 · 2017: 1 · 2015: 1 · 2014: 1 · 2013: 6 · 2012: 3
FWCI
3.66
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Adelaide Research & Scholarship (AR&S) (University of Adelaide) (submittedVersion)
Topics
  • Health Sciences › Medicine › Genetics › Hemoglobinopathies and Related Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Porphyrin Metabolism and Disorders
Keywords
5-aminolevulinate synthase, erythroid-specific gene expression, promoter regulation, GATA-1, CACCC box, EKLF, Sp1, TATA box, transcription factor binding sites, enhancers, heme biosynthesis, erythroid cells
MeSH terms
5-Aminolevulinate Synthetase, Binding Sites, Leukemia, Erythroblastic, Acute, Humans, Promoter Regions, Genetic, Transcription Factors, Transcription, Genetic, Tumor Cells, Cultured, Gene Expression Regulation, Enzymologic, Mutagenesis, Site-Directed, Genes, Reporter

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