Abstract
Formamide gel electrophoresis separates the mRNA fraction from reticulocyte polyribosomes of adult humans into two major RNA species with migratory rates identical to those of the alpha- and beta-globin mRNAs of the rabbit. That these two RNAs of human origin are the globin mRNAs is further supported by the deficiency of the presumed beta mRNA in reticulocyte polyribosomes of fetuses and premature infants, whose cells make gamma chains in preference to beta chains. The globin mRNAs of reticulocyte polyribosomes from patients with hematological disorders were estimated by scanning the stained formamide gels. In contrast to individuals with either hemolytic anemia without hemoglobinopathy or sickle cell anemia who had beta mRNA to alpha mRNA ratios of approximately one, a patient with Hb S-beta-thalassemia had a ratio of beta mRNA to alpha mRNA of 0.75 while two subjects with homozygous beta-thalassemia had severe deficiencies of beta mRNA. Conversely, a patient with alpha-thalassemia (Hb H disease) had a ratio of beta mRNA to alpha mRNA on reticulocyte polyribosomes of 6. These data provide further evidence of a quantitative deficiency of chain-specific globin mRNA in patients with the thalassemia syndromes.
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- Aviv H., Boime I., Leder P. Protein synthesis directed by encephalomyocarditis virus RNA: properties of a transfer RNA-dependent system. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2303–2307. doi: 10.1073/pnas.68.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bank A., Terada M., Metafora S., Dow L., Marks P. A. In vitro synthesis of DNA components of human genes for globins. Nat New Biol. 1972 Feb 9;235(58):167–169. doi: 10.1038/newbio235167a0. [DOI] [PubMed] [Google Scholar]
- Benz E. J., Jr, Forget B. G. Defect in messenger RNA for human hemoglobin synthesis in beta thalassemia. J Clin Invest. 1971 Dec;50(12):2755–2760. doi: 10.1172/JCI106778. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Charache S., Conley C. L., Doeblin T. D., Bartalos M. Thalassemia in black americans. Ann N Y Acad Sci. 1974;232(0):125–134. doi: 10.1111/j.1749-6632.1974.tb20577.x. [DOI] [PubMed] [Google Scholar]
- Clegg J. B., Naughton M. A., Weatherball D. J. Abnormal human haemoglobins. Separation and characterization of the alpha and beta chains by chromatography, and the determination of two new variants, hb Chesapeak and hb J (Bangkok). J Mol Biol. 1966 Aug;19(1):91–108. doi: 10.1016/s0022-2836(66)80052-9. [DOI] [PubMed] [Google Scholar]
- Clegg J. B., Weatherall D. J., Contopolou-Griva I., Caroutsos K., Poungouras P., Tsevrenis H. Haemoglobin Icaria, a new chain-termination mutant with causes alpha thalassaemia. Nature. 1974 Sep 20;251(5472):245–247. doi: 10.1038/251245a0. [DOI] [PubMed] [Google Scholar]
- Clegg J. B., Weatherall D. J. Haemoglobin synthesis in alpha-thalassaemia (haemoglobin H disease). Nature. 1967 Sep 16;215(5107):1241–1243. doi: 10.1038/2151241a0. [DOI] [PubMed] [Google Scholar]
- Clegg J. B., Weatherall D. J., Milner P. F. Haemoglobin Constant Spring--a chain termination mutant? Nature. 1971 Dec 10;234(5328):337–340. doi: 10.1038/234337a0. [DOI] [PubMed] [Google Scholar]
- Conconi F., Rowley P. T., Del Senno L., Pontremoli S., Volpato S. Induction of -globin synthesis in the -thalassaemia of Ferrara. Nat New Biol. 1972 Jul 19;238(81):83–87. doi: 10.1038/newbio238083a0. [DOI] [PubMed] [Google Scholar]
- Dahlberg A. E., Dingman C. W., Peacock A. C. Electrophoretic characterization of bacterial polyribosomes in agarose-acrylamide composite gels. J Mol Biol. 1969 Apr 14;41(1):139–147. doi: 10.1016/0022-2836(69)90131-4. [DOI] [PubMed] [Google Scholar]
- Evans M. J., Lingrel J. B. Hemoglobin messenger ribonucleic acid. Synthesis of 9S and ribosomal ribonucleic acid during erythroid cell development. Biochemistry. 1969 Jul;8(7):3000–3005. doi: 10.1021/bi00835a048. [DOI] [PubMed] [Google Scholar]
- Forget B. G., Benz E. J., Jr, Skoultchi A., Baglioni C., Housman D. Absence of messenger RNA for beta globin chain in beta(0) thalassaemia. Nature. 1974 Feb 8;247(5440):379–381. doi: 10.1038/247379a0. [DOI] [PubMed] [Google Scholar]
- Gould H. J., Hamlyn P. H. The molecular weight of rabbit globin messenger RNA's. FEBS Lett. 1973 Mar 15;30(3):301–304. doi: 10.1016/0014-5793(73)80674-x. [DOI] [PubMed] [Google Scholar]
- Grossbard E., Terada M., Dow L. W., Bank A. Decreased globin messenger RNA activity associated with polyribosomes in thalassaemia. Nat New Biol. 1973 Feb 14;241(111):209–211. doi: 10.1038/newbio241209a0. [DOI] [PubMed] [Google Scholar]
- HUISMAN T. H., PUNT K., SCHAAD J. D. Thalassemia minor associated with hemoglobin-B2 heterozygosity. A family report. Blood. 1961 Jun;17:747–757. [PubMed] [Google Scholar]
- Heywood D., Karon M., Weissman S. Asymmetrical incorporation of amino acids in the alpha and beta chains of hemoglobin synthesized by thalassemic reticulocytes. J Lab Clin Med. 1965 Sep;66(3):476–482. [PubMed] [Google Scholar]
- Hollenberg M. D., Kaback M. M., Kazazian H. H., Jr Adult hemoglobin synthesis by reticulocytes from the human fetus at midtrimester. Science. 1971 Nov 12;174(4010):698–702. doi: 10.1126/science.174.4010.698. [DOI] [PubMed] [Google Scholar]
- Housman D., Forget B. G., Skoultchi A., Benz E. J., Jr Quantitative deficiency of chain-specific globin messenger ribonucleic acids in the thalassemia syndromes. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1809–1813. doi: 10.1073/pnas.70.6.1809. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kacian D. L., Gambino R., Dow L. W., Grossbard E., Natta C., Ramirez F., Spiegelman S., Marks P. A., Bank A. Decreased globin messenger RNA in thalassemia detected by molecular hybridization. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1886–1890. doi: 10.1073/pnas.70.6.1886. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kazazian H. H., Jr Separation of alpha- and beta-globin messenger RNAs. Nat New Biol. 1972 Aug 9;238(84):166–169. doi: 10.1038/newbio238166a0. [DOI] [PubMed] [Google Scholar]
- Kazazian H. H., Jr, Snyder P. G., Cheng T. C. Separation of alpha- and beta-globin messenger RNAs by formamide gel electrophoresis. Biochem Biophys Res Commun. 1974 Aug 5;59(3):1053–1061. doi: 10.1016/s0006-291x(74)80086-0. [DOI] [PubMed] [Google Scholar]
- Kazazian H. H., Jr, Woodhead A. P. Hemoglobin A synthesis in the developing fetus. N Engl J Med. 1973 Jul 12;289(2):58–62. doi: 10.1056/NEJM197307122890202. [DOI] [PubMed] [Google Scholar]
- Labrie F. Isolation of an RNA with the properties of haemoglobin messenger. Nature. 1969 Mar 29;221(5187):1217–1222. doi: 10.1038/2211217a0. [DOI] [PubMed] [Google Scholar]
- Mathews M., Korner A. Mammalian cell-free protein synthesis directed by viral ribonucleic acid. Eur J Biochem. 1970 Dec;17(2):328–338. doi: 10.1111/j.1432-1033.1970.tb01170.x. [DOI] [PubMed] [Google Scholar]
- Morrison M. R., Brinkley S. A., Gorski J., Lingrel J. B. The separation and identification of alpha- and beta-globin messenger ribonucleic acids. J Biol Chem. 1974 Aug 25;249(16):5290–5295. [PubMed] [Google Scholar]
- Natta C., Banks J., Niazi G., Marks P. A., Bank A. Decreased beta globin mRNA activity in bone marrow cells in homozygous and heterozygous beta thalassaemia. Nat New Biol. 1973 Aug 29;244(139):280–281. doi: 10.1038/newbio244280a0. [DOI] [PubMed] [Google Scholar]
- Nienhuis A. W., Anderson W. F. Isolation and translation of hemoglobin messenger RNA from thalassemia, sickle cell anemia, and normal human reticulocytes. J Clin Invest. 1971 Nov;50(11):2458–2460. doi: 10.1172/JCI106745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nienhuis A. W., Canfield P. H., Anderson W. F. Hemoglobin messenger RNA from human bone marrow. Isolation and translation in homozygous and heterozygous beta-thalassemia. J Clin Invest. 1973 Jul;52(7):1735–1745. doi: 10.1172/JCI107355. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ottolenghi S., Lanyon W. G., Paul J., Williamson R., Weatherall D. J., Clegg J. B., Pritchard J., Pootrakul S., Boon W. H. The severe form of alpha thalassaemia is caused by a haemoglobin gene deletion. Nature. 1974 Oct 4;251(5474):389–392. doi: 10.1038/251389a0. [DOI] [PubMed] [Google Scholar]
- PEARSON H. A., MOORE M. M. HUMAN HEMOGLOBIN GENE LINKAGE: REPORT OF A FAMILY WITH HEMOGLOBIN B2, HEMOGLOBIN S, AND BETA THALASSEMIA, INCLUDING A PROBABLE CROSSOVER BETWEEN THALASSEMIA AND DELTA LOCI. Am J Hum Genet. 1965 Mar;17:125–132. [PMC free article] [PubMed] [Google Scholar]
- Ross J., Aviv H., Scolnick E., Leder P. In vitro synthesis of DNA complementary to purified rabbit globin mRNA (RNA-dependent DNA polymerase-reticulocyte-hemoglobin-density gradient centrifugation-oligo(dT) primer). Proc Natl Acad Sci U S A. 1972 Jan;69(1):264–268. doi: 10.1073/pnas.69.1.264. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Staynov D. Z., Pinder J. C., Gratzer W. B. Molecular weight determination of nucleic acids by gel electrophoresis in non-aqueous solution. Nat New Biol. 1972 Jan 26;235(56):108–110. doi: 10.1038/newbio235108a0. [DOI] [PubMed] [Google Scholar]
- Taylor J. M., Dozy A., Kan Y. W., Varmus H. E., Lie-Injo L. E., Ganesan J., Todd D. Genetic lesion in homozygous alpha thalassaemia (hydrops fetalis). Nature. 1974 Oct 4;251(5474):392–393. doi: 10.1038/251392a0. [DOI] [PubMed] [Google Scholar]
- Thompson R. B., Odom J., Bell W. N. Hb-S, beta thalassemia and Hb-A-2 (B-2) in a family with evidence of a crossover between beta and delta loci. Acta Genet Stat Med. 1965;15(3):371–377. doi: 10.1159/000151926. [DOI] [PubMed] [Google Scholar]
- Verma I. M., Temple G. F., Fan H., Baltimore D. In vitro synthesis of DNA complementary to rabbit reticulocyte 10S RNA. Nat New Biol. 1972 Feb 9;235(58):163–167. doi: 10.1038/newbio235163a0. [DOI] [PubMed] [Google Scholar]
- Weatherall D. J., Clegg J. B., Naughton M. A. Globin synthesis in thalassaemia: an in vitro study. Nature. 1965 Dec 11;208(5015):1061–1065. doi: 10.1038/2081061a0. [DOI] [PubMed] [Google Scholar]
- Wolf J. L., Mason R. G., Honig G. R. Regulation of hemoglobin beta-chain synthesis in bone marrow erythroid cells by alpha chains. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3405–3409. doi: 10.1073/pnas.70.12.3405. [DOI] [PMC free article] [PubMed] [Google Scholar]