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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1988 Nov;85(21):8081–8085. doi: 10.1073/pnas.85.21.8081

Asynchronous DNA replication within the human beta-globin gene locus.

E Epner 1, W C Forrester 1, M Groudine 1
PMCID: PMC282358  PMID: 3186709

Abstract

The timing of DNA replication of the human beta-globin gene locus has been studied by blot hybridization of newly synthesized BrdUrd-substituted DNA from cells in different stages of the S phase. Using probes that span greater than 120 kilobases across the human beta-globin gene locus, we show that the majority of this domain replicates in early S phase in the human erythroleukemia cell line K562 and in middle-to-late S phase in the lymphoid cell line Manca. However, in K562 cells three small regions display a strikingly different replication pattern than adjacent sequences. These islands, located in the inter-gamma-globin gene region and approximately 20 kilobases 5' to the epsilon-globin gene and 20 kilobases 3' to the beta-globin gene, replicate later and throughout S phase. A similar area is also present in the alpha-globin gene region in K562 cells. We suggest that these regions may represent sites of termination of replication forks.

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Selected References

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  1. Bell G. I., Gerhard D. S., Fong N. M., Sanchez-Pescador R., Rall L. B. Isolation of the human insulin-like growth factor genes: insulin-like growth factor II and insulin genes are contiguous. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6450–6454. doi: 10.1073/pnas.82.19.6450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bentley D. L., Groudine M. Novel promoter upstream of the human c-myc gene and regulation of c-myc expression in B-cell lymphomas. Mol Cell Biol. 1986 Oct;6(10):3481–3489. doi: 10.1128/mcb.6.10.3481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown E. H., Iqbal M. A., Stuart S., Hatton K. S., Valinsky J., Schildkraut C. L. Rate of replication of the murine immunoglobulin heavy-chain locus: evidence that the region is part of a single replicon. Mol Cell Biol. 1987 Jan;7(1):450–457. doi: 10.1128/mcb.7.1.450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chang E. H., Gonda M. A., Ellis R. W., Scolnick E. M., Lowy D. R. Human genome contains four genes homologous to transforming genes of Harvey and Kirsten murine sarcoma viruses. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4848–4852. doi: 10.1073/pnas.79.16.4848. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Collins F. S., Weissman S. M. The molecular genetics of human hemoglobin. Prog Nucleic Acid Res Mol Biol. 1984;31:315–462. doi: 10.1016/s0079-6603(08)60382-7. [DOI] [PubMed] [Google Scholar]
  6. Edenberg H. J., Huberman J. A. Eukaryotic chromosome replication. Annu Rev Genet. 1975;9:245–284. doi: 10.1146/annurev.ge.09.120175.001333. [DOI] [PubMed] [Google Scholar]
  7. Epner E., Rifkind R. A., Marks P. A. Replication of alpha and beta globin DNA sequences occurs during early S phase in murine erythroleukemia cells. Proc Natl Acad Sci U S A. 1981 May;78(5):3058–3062. doi: 10.1073/pnas.78.5.3058. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Forrester W. C., Takegawa S., Papayannopoulou T., Stamatoyannopoulos G., Groudine M. Evidence for a locus activation region: the formation of developmentally stable hypersensitive sites in globin-expressing hybrids. Nucleic Acids Res. 1987 Dec 23;15(24):10159–10177. doi: 10.1093/nar/15.24.10159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Forrester W. C., Thompson C., Elder J. T., Groudine M. A developmentally stable chromatin structure in the human beta-globin gene cluster. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1359–1363. doi: 10.1073/pnas.83.5.1359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Furst A., Brown E. H., Braunstein J. D., Schildkraut C. L. alpha-Globulin sequences are located in a region of early-replicating DNA in murine erythroleukemia cells. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1023–1027. doi: 10.1073/pnas.78.2.1023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gerhard D. S., Jones C., Morse H. G., Handelin B., Weeks V., Housman D. Analysis of human chromosome 11 by somatic cell genetics: reexamination of derivatives of human-hamster cell line J1. Somat Cell Mol Genet. 1987 Jul;13(4):293–304. doi: 10.1007/BF01534923. [DOI] [PubMed] [Google Scholar]
  12. Germino J., Bastia D. Termination of DNA replication in vitro at a sequence-specific replication terminus. Cell. 1981 Mar;23(3):681–687. doi: 10.1016/0092-8674(81)90431-1. [DOI] [PubMed] [Google Scholar]
  13. Goldman M. A., Holmquist G. P., Gray M. C., Caston L. A., Nag A. Replication timing of genes and middle repetitive sequences. Science. 1984 May 18;224(4650):686–692. doi: 10.1126/science.6719109. [DOI] [PubMed] [Google Scholar]
  14. Grosveld F., van Assendelft G. B., Greaves D. R., Kollias G. Position-independent, high-level expression of the human beta-globin gene in transgenic mice. Cell. 1987 Dec 24;51(6):975–985. doi: 10.1016/0092-8674(87)90584-8. [DOI] [PubMed] [Google Scholar]
  15. Groudine M., Kohwi-Shigematsu T., Gelinas R., Stamatoyannopoulos G., Papayannopoulou T. Human fetal to adult hemoglobin switching: changes in chromatin structure of the beta-globin gene locus. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7551–7555. doi: 10.1073/pnas.80.24.7551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Groudine M., Weintraub H. Propagation of globin DNAase I-hypersensitive sites in absence of factors required for induction: a possible mechanism for determination. Cell. 1982 Aug;30(1):131–139. doi: 10.1016/0092-8674(82)90019-8. [DOI] [PubMed] [Google Scholar]
  17. Henthorn P. S., Mager D. L., Huisman T. H., Smithies O. A gene deletion ending within a complex array of repeated sequences 3' to the human beta-globin gene cluster. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5194–5198. doi: 10.1073/pnas.83.14.5194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Holmquist G. P. Role of replication time in the control of tissue-specific gene expression. Am J Hum Genet. 1987 Feb;40(2):151–173. [PMC free article] [PubMed] [Google Scholar]
  19. Holmquist G., Gray M., Porter T., Jordan J. Characterization of Giemsa dark- and light-band DNA. Cell. 1982 Nov;31(1):121–129. doi: 10.1016/0092-8674(82)90411-1. [DOI] [PubMed] [Google Scholar]
  20. Kioussis D., Vanin E., deLange T., Flavell R. A., Grosveld F. G. Beta-globin gene inactivation by DNA translocation in gamma beta-thalassaemia. Nature. 1983 Dec 15;306(5944):662–666. doi: 10.1038/306662a0. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Nishikori M., Hansen H., Jhanwar S., Fried J., Sordillo P., Koziner B., Lloyd K., Clarkson B. Establishment of a near-tetraploid B-cell lymphoma line with duplication of the 8;14 translocation. Cancer Genet Cytogenet. 1984 May;12(1):39–50. doi: 10.1016/0165-4608(84)90006-2. [DOI] [PubMed] [Google Scholar]
  23. Quan F., Korneluk R. G., Tropak M. B., Gravel R. A. Isolation and characterization of the human catalase gene. Nucleic Acids Res. 1986 Jul 11;14(13):5321–5335. doi: 10.1093/nar/14.13.5321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ravetch J. V., Siebenlist U., Korsmeyer S., Waldmann T., Leder P. Structure of the human immunoglobulin mu locus: characterization of embryonic and rearranged J and D genes. Cell. 1981 Dec;27(3 Pt 2):583–591. doi: 10.1016/0092-8674(81)90400-1. [DOI] [PubMed] [Google Scholar]
  25. Smithies O. The control of globin and other eukaryotic genes. J Cell Physiol Suppl. 1982;1:137–143. doi: 10.1002/jcp.1041130421. [DOI] [PubMed] [Google Scholar]
  26. Taramelli R., Kioussis D., Vanin E., Bartram K., Groffen J., Hurst J., Grosveld F. G. Gamma delta beta-thalassaemias 1 and 2 are the result of a 100 kbp deletion in the human beta-globin cluster. Nucleic Acids Res. 1986 Sep 11;14(17):7017–7029. doi: 10.1093/nar/14.17.7017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Thompson C. B., Challoner P. B., Neiman P. E., Groudine M. Levels of c-myc oncogene mRNA are invariant throughout the cell cycle. 1985 Mar 28-Apr 3Nature. 314(6009):363–366. doi: 10.1038/314363a0. [DOI] [PubMed] [Google Scholar]
  28. Tuan D., Feingold E., Newman M., Weissman S. M., Forget B. G. Different 3' end points of deletions causing delta beta-thalassemia and hereditary persistence of fetal hemoglobin: implications for the control of gamma-globin gene expression in man. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6937–6941. doi: 10.1073/pnas.80.22.6937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tuan D., Solomon W., Li Q., London I. M. The "beta-like-globin" gene domain in human erythroid cells. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6384–6388. doi: 10.1073/pnas.82.19.6384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Vasicek T. J., McDevitt B. E., Freeman M. W., Fennick B. J., Hendy G. N., Potts J. T., Jr, Rich A., Kronenberg H. M. Nucleotide sequence of the human parathyroid hormone gene. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2127–2131. doi: 10.1073/pnas.80.8.2127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weaver D. T., Fields-Berry S. C., DePamphilis M. L. The termination region for SV40 DNA replication directs the mode of separation for the two sibling molecules. Cell. 1985 Jun;41(2):565–575. doi: 10.1016/s0092-8674(85)80029-5. [DOI] [PubMed] [Google Scholar]
  32. Weintraub H. Assembly of an active chromatin structure during replication. Nucleic Acids Res. 1979 Oct 10;7(3):781–792. doi: 10.1093/nar/7.3.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Weintraub H., Flint S. J., Leffak I. M., Groudine M., Grainger R. M. The generation and propagation of variegated chromosome structures. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):401–407. doi: 10.1101/sqb.1978.042.01.042. [DOI] [PubMed] [Google Scholar]
  34. Yagi M., Gelinas R., Elder J. T., Peretz M., Papayannopoulou T., Stamatoyannopoulos G., Groudine M. Chromatin structure and developmental expression of the human alpha-globin cluster. Mol Cell Biol. 1986 Apr;6(4):1108–1116. doi: 10.1128/mcb.6.4.1108. [DOI] [PMC free article] [PubMed] [Google Scholar]

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