Abstract
We studied the abundance and structure of globin mRNAs present in K562 cells both before and after induction of hemoglobin synthesis by hemin. In vitro translation of poly(A)+ RNA from K562 cells generated protein products corresponding to alpha, A gamma-, G gamma-, epsilon-, and zeta-globin mRNAs. Individual globin mRNAs increased 1.5- to 3-fold after induction. Similar results were obtained by measuring steady-state mRNA concentrations of induced and uninduced cells by using S1 nuclease mapping. Globin gene transcripts were correctly initiated and processed. In addition, S1 nuclease analysis revealed the presence of delta-globin mRNA in both control and induced cells. A small percentage of delta-globin transcripts appeared to be initiated upstream from the normal initiation site. beta-Globin mRNA was not detected in any studies. The results (i) suggest that hemin induction of K562 cells is mediated at a transcriptional level and (ii) reveal the dissociation of delta- and beta-globin gene expression in K562 cells compared with normal erythroid cells.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adams S. L., Sobel M. E., Howard B. H., Olden K., Yamada K. M., de Crombrugghe B., Pastan I. Levels of translatable mRNAs for cell surface protein, collagen precursors, and two membrane proteins are altered in Rous sarcoma virus-transformed chick embryo fibroblasts. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3399–3403. doi: 10.1073/pnas.74.8.3399. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Alan M., Grindlay G. J., Stefani L., Paul J. Epsilon globin gene transcripts originating upstream of the mRNA cap site in K562 cells and normal human embryos. Nucleic Acids Res. 1982 Sep 11;10(17):5133–5147. doi: 10.1093/nar/10.17.5133. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Alter B. P., Goff S. C. Electrophoretic separation of human embryonic globin demonstrates "alpha-thalassemia" in human leukemia cell line K562. Biochem Biophys Res Commun. 1980 Jun 16;94(3):843–848. doi: 10.1016/0006-291x(80)91311-x. [DOI] [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., Mears J. G., Ramirez F. Disorders of human hemoglobin. Science. 1980 Feb 1;207(4430):486–493. doi: 10.1126/science.7352255. [DOI] [PubMed] [Google Scholar]
- Benz E. J., Jr, Murnane M. J., Tonkonow B. L., Berman B. W., Mazur E. M., Cavallesco C., Jenko T., Snyder E. L., Forget B. G., Hoffman R. Embryonic-fetal erythroid characteristics of a human leukemic cell line. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3509–3513. doi: 10.1073/pnas.77.6.3509. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Charnay P., Maniatis T. Transcriptional regulation of globin gene expression in the human erythroid cell line K562. Science. 1983 Jun 17;220(4603):1281–1283. doi: 10.1126/science.6574602. [DOI] [PubMed] [Google Scholar]
- Cioe L., McNab A., Hubbell H. R., Meo P., Curtis P., Rovera G. Differential expression of the globin genes in human leukemia K562(S) cells induced to differentiate by hemin or butyric acid. Cancer Res. 1981 Jan;41(1):237–243. [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]
- Dean A., Erard F., Schneider A. P., Schechter A. N. Induction of hemoglobin accumulation in human K562 cells by hemin is reversible. Science. 1981 Apr 24;212(4493):459–461. doi: 10.1126/science.6163216. [DOI] [PubMed] [Google Scholar]
- Erard F., Dean A., Schechter A. N. Inhibitors of cell division reversibly modify hemoglobin concentration in human erythroleukemia K562 cells. Blood. 1981 Dec;58(6):1236–1239. [PubMed] [Google Scholar]
- Groudine M., Weintraub H. Activation of globin genes during chicken development. Cell. 1981 May;24(2):393–401. doi: 10.1016/0092-8674(81)90329-9. [DOI] [PubMed] [Google Scholar]
- Humphries R. K., Ley T., Turner P., Moulton A. D., Nienhuis A. W. Differences in human alpha-, beta- and delta-globin gene expression in monkey kidney cells. Cell. 1982 Aug;30(1):173–183. doi: 10.1016/0092-8674(82)90023-x. [DOI] [PubMed] [Google Scholar]
- Lawn R. M., Efstratiadis A., O'Connell C., Maniatis T. The nucleotide sequence of the human beta-globin gene. Cell. 1980 Oct;21(3):647–651. doi: 10.1016/0092-8674(80)90428-6. [DOI] [PubMed] [Google Scholar]
- Ley T. J., Anagnou N. P., Pepe G., Nienhuis A. W. RNA processing errors in patients with beta-thalassemia. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4775–4779. doi: 10.1073/pnas.79.15.4775. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ley T. J., Nienhuis A. W. A weak upstream promoter gives rise to long human beta-globin RNA molecules. Biochem Biophys Res Commun. 1983 May 16;112(3):1041–1048. doi: 10.1016/0006-291x(83)91723-0. [DOI] [PubMed] [Google Scholar]
- Lozzio C. B., Lozzio B. B. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood. 1975 Mar;45(3):321–334. [PubMed] [Google Scholar]
- Ricca G. A., Taylor J. M., Kalinyak J. E. Simple rapid method for the synthesis of radioactively labeled cDNA hybridization probes utilizing bacteriophage M13mp7. Proc Natl Acad Sci U S A. 1982 Feb;79(3):724–728. doi: 10.1073/pnas.79.3.724. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rutherford T. R., Clegg J. B., Weatherall D. J. K562 human leukaemic cells synthesise embryonic haemoglobin in response to haemin. Nature. 1979 Jul 12;280(5718):164–165. doi: 10.1038/280164a0. [DOI] [PubMed] [Google Scholar]
- Rutherford T., Clegg J. B., Higgs D. R., Jones R. W., Thompson J., Weatherall D. J. Embryonic erythroid differentiation in the human leukemic cell line K562. Proc Natl Acad Sci U S A. 1981 Jan;78(1):348–352. doi: 10.1073/pnas.78.1.348. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spritz R. A., DeRiel J. K., Forget B. G., Weissman S. M. Complete nucleotide sequence of the human delta-globin gene. Cell. 1980 Oct;21(3):639–646. doi: 10.1016/0092-8674(80)90427-4. [DOI] [PubMed] [Google Scholar]
- Tonkonow B. L., Hoffman R., Burger D., Elder J. T., Mazur E. M., Murnane M. J., Benz E. J., Jr Differing responses of globin and glycophorin gene expression to hemin in the human leukemia cell line K562. Blood. 1982 Apr;59(4):738–746. [PubMed] [Google Scholar]
- Tsapis A., Hinard N., Testa U., Dubart A., Vainchenker W., Rouyer-Fessard P., Beuzard Y., Rosa J. Globin-chain affinity chromatography on Sepharose-haptoglobin: a new method of study of hemoglobin synthesis in reticulocytes, in bone marrow and in colonies of erythroid precursors. Eur J Biochem. 1980 Dec;112(3):513–519. doi: 10.1111/j.1432-1033.1980.tb06114.x. [DOI] [PubMed] [Google Scholar]
- Wood W. G., Old J. M., Roberts A. V., Clegg J. B., Weatherall D. J. Human globin gene expression: control of beta, delta and delta beta chain production. Cell. 1978 Oct;15(2):437–446. doi: 10.1016/0092-8674(78)90013-2. [DOI] [PubMed] [Google Scholar]