Abstract
Purified alpha and beta globin complementary DNAs (cDNAs) have been separated from total radioactively labeled human globin cDNA using mRNA purified from liver of a hydrops fetalis (alpha thalassemia). The beta cDNA hybridizes to the hydrops fetalis mRNA while the alpha cDNA remains single-stranded. the purified alpha and beta cDNAs were assayed for their purity by their hybridization to mRNA prepared from reticulocytes of nonthalassemia, alpha thalassemia, and beta thalassemia subjects. The results indicate that the separated cDNAs are selective in hybridization to alpha or beta globin mRNAs, respectively. The previously reported deficiency of globin mRNA in thalassemia cells has been confirmed with these purified cDNAs. The purified alpha and beta cDNAs were hybridized to cellular DNA to non-thalassemia, beta+ thalassemia, and hydrops fetalis (alpha thalassemia) DNA. The alpha cDNA hybridized to hydrops fetalis liver DNA to a much lower extent that beta cDNA, confirming the previously reported deletion of alpha globin genes in hydrops fetalis. By contrast, both the alpha and beta DNA probes hybridized to the same extent to spleen DNA from non-thalassemia and from beta+ thalassemia patients. Between two and five globin genes in non-thalassemia and beta+ thalassemia DNA hybridize to beta cDNA and one to five to alpha cDNA. These studies indicate that in beta+ thalassemia, there is no detectable deletion in beta globin genes. The genetic defect in beta+ thalassemia appears to be due to either repression of transcription of beta globin genes or abnormal processing of beta globin mRNA.
Full text
PDF![1550](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410a/432575/f6f8a3140972/pnas00047-0336.png)
![1551](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410a/432575/678f6fcc0984/pnas00047-0337.png)
![1552](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410a/432575/8fa7ae45c49a/pnas00047-0338.png)
![1553](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410a/432575/f4b0496d2b07/pnas00047-0339.png)
![1554](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/410a/432575/a321c8ad1bcb/pnas00047-0340.png)
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- Britten R. J., Kohne D. E. Repeated sequences in DNA. Hundreds of thousands of copies of DNA sequences have been incorporated into the genomes of higher organisms. Science. 1968 Aug 9;161(3841):529–540. doi: 10.1126/science.161.3841.529. [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]
- Dow L. W., Terada M., Natta C., Metafora S., Grossbard E., Marks P. A., Bank A. Globin synthesis of intact cells and activity of isolated mRNA in -thalassaemia. Nat New Biol. 1973 May 23;243(125):114–116. [PubMed] [Google Scholar]
- Gambino R., Kacian D., O'Donnell J., Ramirez F., Marks P. A., Bank A. A limited number of globin genes in human DNA. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3966–3970. doi: 10.1073/pnas.71.10.3966. [DOI] [PMC free article] [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]
- 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]
- Kacian D. L., Spiegelman S. Use of micrococcal nuclease to monitor hybridization reactions with DNA. Anal Biochem. 1974 Apr;58(2):534–540. doi: 10.1016/0003-2697(74)90221-8. [DOI] [PubMed] [Google Scholar]
- Metafora S., Terada M., Dow L. W., Marks P. A., Bank A. Increased efficiency of exogenous messenger RNA translation in a Krebs ascites cell lysate. Proc Natl Acad Sci U S A. 1972 May;69(5):1299–1303. doi: 10.1073/pnas.69.5.1299. [DOI] [PMC free article] [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]
- 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]
- 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]
- Weatherall D. J., Clegg J. B., Boon W. H. The haemoglobin constitution of infants with the haemoglobin Bart's hydrops foetalis syndrome. Br J Haematol. 1970 Mar;18(3):357–367. doi: 10.1111/j.1365-2141.1970.tb01449.x. [DOI] [PubMed] [Google Scholar]