Transcriptional Regulation of the Human Erythroid 5-Aminolevulinate Synthase Gene
Open accessAbstract
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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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].”
From Aminolevulinic Acid (ALA) as a Prodrug in Photodynamic Therapy of Cancer (2011) · cites this paper as [37]
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Fratz‐Berilla, E. J. (2014) · Digital Commons - University of South Florida (University of South Florida)
- Authors
- Surinya, K. H., Cox, T. C., & May, B. K.
- Affiliations
- The University of Adelaide (AU)
- Corresponding author
- May, Brian K.
- ORCID
- Cox, Timothy C.
- 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
- hybrid
- doi.org
- Licence
- 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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