HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Surinya, K. H. Articles by May, B. K. Search for Related Content PubMed PubMed Citation Articles by Surinya, K. H. Articles by May, B. K. Social Bookmarking                      What's this?

Volume 272, Number 42, Issue of October 17, 1997 pp. 26585-26594
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Transcriptional Regulation of the Human Erythroid 5-Aminolevulinate Synthase Gene
IDENTIFICATION OF PROMOTER ELEMENTS AND ROLE OF REGULATORY PROTEINS

(Received for publication, February 10, 1997, and in revised form, July 7, 1997)

Katharina H. Surinya , Timothy C. Cox and Brian K. May

From the Department of Biochemistry, University of Adelaide, South Australia, 5005 Australia and the  Department of Genetics, University of Adelaide, South Australia, 5005 Australia

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.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				C. R. Scherzer, J. A. Grass, Z. Liao, I. Pepivani, B. Zheng, A. C. Eklund, P. A. Ney, J. Ng, M. McGoldrick, B. Mollenhauer, et al. GATA transcription factors directly regulate the Parkinson's disease-linked gene {alpha}-synuclein PNAS, August 5, 2008; 105(31): 10907 - 10912. [Abstract] [Full Text] [PDF]

				W. J. Bakker, T. B. van Dijk, M. Parren-van Amelsvoort, A. Kolbus, K. Yamamoto, P. Steinlein, R. G. W. Verhaak, T. W. Mak, H. Beug, B. Lowenberg, et al. Differential Regulation of Foxo3a Target Genes in Erythropoiesis Mol. Cell. Biol., May 15, 2007; 27(10): 3839 - 3854. [Abstract] [Full Text] [PDF]

				Y.-C. Shyu, T.-L. Lee, S.-C. Wen, H. Chen, W.-Y. Hsiao, X. Chen, J. Hwang, and C.-K. J. Shen Subcellular Transport of EKLF and Switch-On of Murine Adult {beta}maj Globin Gene Transcription Mol. Cell. Biol., March 15, 2007; 27(6): 2309 - 2323. [Abstract] [Full Text] [PDF]

				O. Nakajima, S. Okano, H. Harada, T. Kusaka, X. Gao, T. Hosoya, N. Suzuki, S. Takahashi, and M. Yamamoto Transgenic rescue of erythroid 5-aminolevulinate synthase-deficient mice results in the formation of ring sideroblasts and siderocytes Genes Cells, June 1, 2006; 11(6): 685 - 700. [Abstract] [Full Text] [PDF]

				D. Hodge, E. Coghill, J. Keys, T. Maguire, B. Hartmann, A. McDowall, M. Weiss, S. Grimmond, and A. Perkins A global role for EKLF in definitive and primitive erythropoiesis Blood, April 15, 2006; 107(8): 3359 - 3370. [Abstract] [Full Text] [PDF]

				N. Ueki, L. Zhang, and M. J. Hayman Ski Negatively Regulates Erythroid Differentiation through Its Interaction with GATA1 Mol. Cell. Biol., December 1, 2004; 24(23): 10118 - 10125. [Abstract] [Full Text] [PDF]

				S. Bekri, A. May, P. D. Cotter, A. I. Al-Sabah, X. Guo, G. S. Masters, and D. F. Bishop A promoter mutation in the erythroid-specific 5-aminolevulinate synthase (ALAS2) gene causes X-linked sideroblastic anemia Blood, July 15, 2003; 102(2): 698 - 704. [Abstract] [Full Text] [PDF]

				S. E. Lyons, N. D. Lawson, L. Lei, P. E. Bennett, B. M. Weinstein, and P. P. Liu A nonsense mutation in zebrafish gata1 causes the bloodless phenotype in vlad tepes PNAS, April 16, 2002; 99(8): 5454 - 5459. [Abstract] [Full Text] [PDF]

				E. Coghill, S. Eccleston, V. Fox, L. Cerruti, C. Brown, J. Cunningham, S. Jane, and A. Perkins Erythroid Kruppel-like factor (EKLF) coordinates erythroid cell proliferation and hemoglobinization in cell lines derived from EKLF null mice Blood, March 15, 2001; 97(6): 1861 - 1868. [Abstract] [Full Text] [PDF]

				A. C. Perkins, K. R. Peterson, G. Stamatoyannopoulos, H. E. Witkowska, and S. H. Orkin Fetal expression of a human Agamma globin transgene rescues globin chain imbalance but not hemolysis in EKLF null mouse embryos Blood, March 1, 2000; 95(5): 1827 - 1833. [Abstract] [Full Text] [PDF]

				H. Asano, X. S. Li, and G. Stamatoyannopoulos FKLF, a Novel Kruppel-Like Factor That Activates Human Embryonic and Fetal beta -Like Globin Genes Mol. Cell. Biol., May 1, 1999; 19(5): 3571 - 3579. [Abstract] [Full Text] [PDF]

				P. G. Gallagher, D. E. Sabatino, M. Romana, A. P. Cline, L. J. Garrett, D. M. Bodine, and B. G. Forget A Human beta -Spectrin Gene Promoter Directs High Level Expression in Erythroid but Not Muscle or Neural Cells J. Biol. Chem., March 5, 1999; 274(10): 6062 - 6073. [Abstract] [Full Text] [PDF]

				A. Spadaccini, P. A. Tilbrook, M. K. Sarna, M. Crossley, J. J. Bieker, and S. P. Klinken Transcription Factor Erythroid Kruppel-like Factor (ELKF) Is Essential for the Erythropoietin-induced Hemoglobin Production but Not for Proliferation, Viability, or Morphological Maturation J. Biol. Chem., September 11, 1998; 273(37): 23793 - 23798. [Abstract] [Full Text] [PDF]

				K. H. Surinya, T. C. Cox, and B. K. May Identification and Characterization of a Conserved Erythroid-specific Enhancer Located in Intron 8 of the Human 5-Aminolevulinate Synthase 2 Gene J. Biol. Chem., July 3, 1998; 273(27): 16798 - 16809. [Abstract] [Full Text] [PDF]