Abstract
Recently, the human gene encoding erythroid-specific delta-aminolevulinate synthase was localized to the chromosomal region Xp21-Xq21, identifying this gene as the logical candidate for the enzymatic defect causing "X-linked" sideroblastic anemia. To investigate this hypothesis, the 11 exonic coding regions of the delta-aminolevulinate synthase gene were amplified and sequenced from a 30-year-old Chinese male with a pyridoxine-responsive form of X-linked sideroblastic anemia. A single T----A transition was found in codon 471 in a highly conserved region of exon 9, resulting in an Ile----Asn substitution. This mutation interrupted contiguous hydrophobic residues and was predicted to transform a region of beta-sheet structure to a random-coil structure. Prokaryotic expression of the normal and mutant cDNAs revealed that the mutant construct expressed low levels of enzymatic activity that required higher concentrations of pyridoxal 5'-phosphate to achieve maximal activation than did the normal enzyme. The amino acid substitution occurred in the exon containing the putative pyridoxal 5'-phosphate binding site and may account for the reduced ability of the cofactor to catalyze the formation of delta-aminolevulinic acid.
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Selected References
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- Aoki Y., Urata G., Wada O., Takaku F. Measurement of delta-aminolevulinic acid synthetase activity in human erythroblasts. J Clin Invest. 1974 May;53(5):1326–1334. doi: 10.1172/JCI107680. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Aronson B. D., Ravnikar P. D., Somerville R. L. Nucleotide sequence of the 2-amino-3-ketobutyrate coenzyme A ligase (kbl) gene of E. coli. Nucleic Acids Res. 1988 Apr 25;16(8):3586–3586. doi: 10.1093/nar/16.8.3586. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bawden M. J., Borthwick I. A., Healy H. M., Morris C. P., May B. K., Elliott W. H. Sequence of human 5-aminolevulinate synthase cDNA. Nucleic Acids Res. 1987 Oct 26;15(20):8563–8563. doi: 10.1093/nar/15.20.8563. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bennett J. M. Classification of the myelodysplastic syndromes. Clin Haematol. 1986 Nov;15(4):909–923. [PubMed] [Google Scholar]
- Bishop D. F., Henderson A. S., Astrin K. H. Human delta-aminolevulinate synthase: assignment of the housekeeping gene to 3p21 and the erythroid-specific gene to the X chromosome. Genomics. 1990 Jun;7(2):207–214. doi: 10.1016/0888-7543(90)90542-3. [DOI] [PubMed] [Google Scholar]
- Bishop D. F., Kitchen H., Wood W. A. Evidence for erythroid and nonerythroid forms of delta-aminolevulinate synthetase. Arch Biochem Biophys. 1981 Feb;206(2):380–391. doi: 10.1016/0003-9861(81)90105-3. [DOI] [PubMed] [Google Scholar]
- Bishop D. F. Two different genes encode delta-aminolevulinate synthase in humans: nucleotide sequences of cDNAs for the housekeeping and erythroid genes. Nucleic Acids Res. 1990 Dec 11;18(23):7187–7188. doi: 10.1093/nar/18.23.7187. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bishop D. F., Wampler D. E., Sgouris J. T., Bonefeld R. J., Anderson D. K., Hawley M. C., Sweeley C. C. Pilot scale purification of alpha-galactosidase A from Cohn fraction IV-1 of human plasma. Biochim Biophys Acta. 1978 May 11;524(1):109–120. doi: 10.1016/0005-2744(78)90109-2. [DOI] [PubMed] [Google Scholar]
- Bishop D. F., Wood W. A. An assay for delta-aminolevulinic acid synthetase based on a specific, semiautomatic determination of picomole quantities of delta-[14C]aminolevulinate. Anal Biochem. 1977 Jun;80(2):466–482. doi: 10.1016/0003-2697(77)90669-8. [DOI] [PubMed] [Google Scholar]
- Borthwick I. A., Srivastava G., Day A. R., Pirola B. A., Snoswell M. A., May B. K., Elliott W. H. Complete nucleotide sequence of hepatic 5-aminolaevulinate synthase precursor. Eur J Biochem. 1985 Aug 1;150(3):481–484. doi: 10.1111/j.1432-1033.1985.tb09047.x. [DOI] [PubMed] [Google Scholar]
- Cox T. C., Bawden M. J., Abraham N. G., Bottomley S. S., May B. K., Baker E., Chen L. Z., Sutherland G. R. Erythroid 5-aminolevulinate synthase is located on the X chromosome. Am J Hum Genet. 1990 Jan;46(1):107–111. [PMC free article] [PubMed] [Google Scholar]
- Cox T. C., Bawden M. J., Martin A., May B. K. Human erythroid 5-aminolevulinate synthase: promoter analysis and identification of an iron-responsive element in the mRNA. EMBO J. 1991 Jul;10(7):1891–1902. doi: 10.1002/j.1460-2075.1991.tb07715.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Drew P. D., Ades I. Z. Regulation of production of embryonic chick liver delta-aminolevulinate synthase: effects of testosterone and of hemin on the mRNA of the enzyme. Biochem Biophys Res Commun. 1986 Oct 15;140(1):81–87. doi: 10.1016/0006-291x(86)91060-0. [DOI] [PubMed] [Google Scholar]
- Hamel B. C., Schretlen E. D. Sideroblastic anaemia. A review of seven paediatric cases. Eur J Pediatr. 1982 Mar;138(2):130–135. doi: 10.1007/BF00441138. [DOI] [PubMed] [Google Scholar]
- Hayashi N., Kurashima Y., Kikuchi G. Mechanism of allylisopropylacetamide-induced increase of -aminolevulinate synthetase in liver mitochondria. V. Mechanism of regulation by hemin of the level of -aminolevulinate synthetase in rat liver mitochondria. Arch Biochem Biophys. 1972 Jan;148(1):10–21. doi: 10.1016/0003-9861(72)90109-9. [DOI] [PubMed] [Google Scholar]
- Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. doi: 10.1016/0378-1119(89)90358-2. [DOI] [PubMed] [Google Scholar]
- Hornberger U., Liebetanz R., Tichy H. V., Drews G. Cloning and sequencing of the hemA gene of Rhodobacter capsulatus and isolation of a delta-aminolevulinic acid-dependent mutant strain. Mol Gen Genet. 1990 May;221(3):371–378. doi: 10.1007/BF00259402. [DOI] [PubMed] [Google Scholar]
- Kushner J. P., Lee G. R., Wintrobe M. M., Cartwright G. E. Idiopathic refractory sideroblastic anemia: clinical and laboratory investigation of 17 patients and review of the literature. Medicine (Baltimore) 1971 May;50(3):139–159. doi: 10.1097/00005792-197105000-00001. [DOI] [PubMed] [Google Scholar]
- MAUZERALL D., GRANICK S. The occurrence and determination of delta-amino-levulinic acid and porphobilinogen in urine. J Biol Chem. 1956 Mar;219(1):435–446. [PubMed] [Google Scholar]
- MacFerrin K. D., Terranova M. P., Schreiber S. L., Verdine G. L. Overproduction and dissection of proteins by the expression-cassette polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1937–1941. doi: 10.1073/pnas.87.5.1937. [DOI] [PMC free article] [PubMed] [Google Scholar]
- May B. K., Bhasker C. R., Bawden M. J., Cox T. C. Molecular regulation of 5-aminolevulinate synthase. Diseases related to heme biosynthesis. Mol Biol Med. 1990 Oct;7(5):405–421. [PubMed] [Google Scholar]
- McClung C. R., Somerville J. E., Guerinot M. L., Chelm B. K. Structure of the Bradyrhizobium japonicum gene hemA encoding 5-aminolevulinic acid synthase. Gene. 1987;54(1):133–139. doi: 10.1016/0378-1119(87)90355-6. [DOI] [PubMed] [Google Scholar]
- Mukherjee J. J., Dekker E. E. 2-Amino-3-ketobutyrate CoA ligase of Escherichia coli: stoichiometry of pyridoxal phosphate binding and location of the pyridoxyllysine peptide in the primary structure of the enzyme. Biochim Biophys Acta. 1990 Jan 19;1037(1):24–29. doi: 10.1016/0167-4838(90)90097-y. [DOI] [PubMed] [Google Scholar]
- Pasanen A., Tenhunen R. Heme synthesis in sideroblastic anaemias. Scand J Haematol Suppl. 1986;45:60–65. doi: 10.1111/j.1600-0609.1986.tb00845.x. [DOI] [PubMed] [Google Scholar]
- Riddle R. D., Yamamoto M., Engel J. D. Expression of delta-aminolevulinate synthase in avian cells: separate genes encode erythroid-specific and nonspecific isozymes. Proc Natl Acad Sci U S A. 1989 Feb;86(3):792–796. doi: 10.1073/pnas.86.3.792. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sassa S., Granick S. Induction of -aminolevulinic acid synthetase in chick embryo liver cells in cluture. Proc Natl Acad Sci U S A. 1970 Oct;67(2):517–522. doi: 10.1073/pnas.67.2.517. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schoenhaut D. S., Curtis P. J. Nucleotide sequence of mouse 5-aminolevulinic acid synthase cDNA and expression of its gene in hepatic and erythroid tissues. Gene. 1986;48(1):55–63. doi: 10.1016/0378-1119(86)90351-3. [DOI] [PubMed] [Google Scholar]
- Schoenhaut D. S., Curtis P. J. Structure of a mouse erythroid 5-aminolevulinate synthase gene and mapping of erythroid-specific DNAse I hypersensitive sites. Nucleic Acids Res. 1989 Sep 12;17(17):7013–7028. doi: 10.1093/nar/17.17.7013. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Srivastava G., Borthwick I. A., Maguire D. J., Elferink C. J., Bawden M. J., Mercer J. F., May B. K. Regulation of 5-aminolevulinate synthase mRNA in different rat tissues. J Biol Chem. 1988 Apr 15;263(11):5202–5209. [PubMed] [Google Scholar]
- Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
- Sutherland G. R., Baker E., Callen D. F., Hyland V. J., May B. K., Bawden M. J., Healy H. M., Borthwick I. A. 5-Aminolevulinate synthase is at 3p21 and thus not the primary defect in X-linked sideroblastic anemia. Am J Hum Genet. 1988 Sep;43(3):331–335. [PMC free article] [PubMed] [Google Scholar]
- Takaku F., Nakao K. Delta-aminolevulinic acid synthetase activity in erythroblasts of patients with sideroblastic anemia. Life Sci II. 1971 Jul 8;10(13):721–726. doi: 10.1016/0024-3205(71)90203-7. [DOI] [PubMed] [Google Scholar]
- Tanaka M., Ota H. A case of primary acquired sideroblastic anemia with deficient -aminolevulinic acid synthetase activity in bone marrow erythroblasts. Tohoku J Exp Med. 1972 Feb;106(2):199–206. doi: 10.1620/tjem.106.199. [DOI] [PubMed] [Google Scholar]
- Urban-Grimal D., Volland C., Garnier T., Dehoux P., Labbe-Bois R. The nucleotide sequence of the HEM1 gene and evidence for a precursor form of the mitochondrial 5-aminolevulinate synthase in Saccharomyces cerevisiae. Eur J Biochem. 1986 May 2;156(3):511–519. doi: 10.1111/j.1432-1033.1986.tb09610.x. [DOI] [PubMed] [Google Scholar]
- Wada O., Sassa S., Takaku F., Yano Y., Uratta G., Nakao K. Different responses of the hepatic and erythropoietic delta-aminolevulinic acid synthetase of mice. Biochim Biophys Acta. 1967 Nov 28;148(2):585–587. doi: 10.1016/0304-4165(67)90165-1. [DOI] [PubMed] [Google Scholar]
- Watanabe N., Hayashi N., Kikuchi G. delta-Aminolevulinate synthase isozymes in the liver and erythroid cells of chicken. Biochem Biophys Res Commun. 1983 Jun 15;113(2):377–383. doi: 10.1016/0006-291x(83)91737-0. [DOI] [PubMed] [Google Scholar]
- Wetmur J. G. DNA probes: applications of the principles of nucleic acid hybridization. Crit Rev Biochem Mol Biol. 1991;26(3-4):227–259. doi: 10.3109/10409239109114069. [DOI] [PubMed] [Google Scholar]
- Woods J. S. Studies on the role of heme in the regulation of delta-aminolevulinic acid synthetase during fetal hepatic development. Mol Pharmacol. 1974 May;10(3):389–397. [PubMed] [Google Scholar]
- Yamamoto M., Fujita H., Watanabe N., Hayashi N., Kikuchi G. An immunochemical study of delta-aminolevulinate synthase and delta-aminolevulinate dehydratase in liver and erythroid cells of rat. Arch Biochem Biophys. 1986 Feb 15;245(1):76–83. doi: 10.1016/0003-9861(86)90191-8. [DOI] [PubMed] [Google Scholar]
- Yamamoto M., Hayashi N., Kikuchi G. Regulation of synthesis and intracellular translocation of delta-aminolevulinate synthase by heme and its relation to the heme saturation of tryptophan pyrrolase in rat liver. Arch Biochem Biophys. 1981 Jul;209(2):451–459. doi: 10.1016/0003-9861(81)90302-7. [DOI] [PubMed] [Google Scholar]
- Yamamoto M., Kure S., Engel J. D., Hiraga K. Structure, turnover, and heme-mediated suppression of the level of mRNA encoding rat liver delta-aminolevulinate synthase. J Biol Chem. 1988 Nov 5;263(31):15973–15979. [PubMed] [Google Scholar]
- Yamamoto M., Yew N. S., Federspiel M., Dodgson J. B., Hayashi N., Engel J. D. Isolation of recombinant cDNAs encoding chicken erythroid delta-aminolevulinate synthase. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3702–3706. doi: 10.1073/pnas.82.11.3702. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yamauchi K., Hayashi N., Kikuchi G. Translocation of delta-aminolevulinate synthase from the cytosol to the mitochondria and its regulation by hemin in the rat liver. J Biol Chem. 1980 Feb 25;255(4):1746–1751. [PubMed] [Google Scholar]