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PMID: 11160905 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Multiple sites of replication initiation in the human beta-globin gene locus.

Nucleic acids research ·Vol. 29 ·No. 3 ·2001-02-01 ·Pages 809-17

Kamath S, Leffak M

Abstract

The cell cycle-dependent, ordered assembly of protein prereplicative complexes suggests that eukaryotic replication origins determine when genomic replication initiates. By comparison, the factors that determine where replication initiates relative to the sites of prereplicative complex formation are not known. In the human globin gene locus previous work showed that replication initiates at a single site 5' to the ss-globin gene when protein synthesis is inhibited by emetine. The present study has examined the pattern of initiation around the genetically defined ss-globin replicator in logarithmically growing HeLa cells, using two PCR-based nascent strand assays. In contrast to the pattern of initiation detected in emetine-treated cells, analysis of the short nascent strands at five positions spanning a 40 kb globin gene region shows that replication initiates at more than one site in non-drug-treated cells. Quantitation of nascent DNA chains confirmed that replication begins at several locations in this domain, including one near the initiation region (IR) identified in emetine-treated cells. However, the abundance of short nascent strands at another initiation site approximately 20 kb upstream is approximately 4-fold as great as that at the IR. The latter site abuts an early S phase replicating fragment previously defined at low resolution in logarithmically dividing cells.

MeSH Terms
DNA/genetics DNA Replication Globins/genetics HeLa Cells Humans Nucleic Acid Hybridization/methods Polymerase Chain Reaction
Chemicals
Globins DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kamath S
Department of Biochemistry and Molecular Biology, Wright State University, Dayton, OH 45435, USA.
Leffak M
References (49)
49 references, click to expand
  1. Initiation of eukaryotic DNA replication: origin unwinding and sequential chromatin association of Cdc45, RPA, and DNA polymerase alpha.
    Mol Cell. 2000 Apr;5(4):617-27 PMID: 10882098
  2. Localization and DNA sequence of a replication origin in the rhodopsin gene locus of Chinese hamster cells.
    J Mol Biol. 1992 Mar 20;224(2):343-58 PMID: 1560457
  3. Molecular cloning and characterization of the human beta-like globin gene cluster.
    Cell. 1980 Apr;19(4):959-72 PMID: 6155216
  4. 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):3058-62 PMID: 6942415
  5. Localized melting and structural changes in the SV40 origin of replication induced by T-antigen.
    EMBO J. 1988 Oct;7(10):3149-58 PMID: 2846276
  6. Asynchronous DNA replication within the human beta-globin gene locus.
    Proc Natl Acad Sci U S A. 1988 Nov;85(21):8081-5 PMID: 3186709
  7. The coordinate replication of the human beta-globin gene domain reflects its transcriptional activity and nuclease hypersensitivity.
    Mol Cell Biol. 1988 Nov;8(11):4958-65 PMID: 2850471
  8. Amplification enhancers and replication origins in the autosomal chorion gene cluster of Drosophila.
    EMBO J. 1989 Mar;8(3):891-901 PMID: 2542026
  9. Activation and repression of a beta-globin gene in cell hybrids is accompanied by a shift in its temporal replication.
    Mol Cell Biol. 1989 Aug;9(8):3524-32 PMID: 2796994
  10. Mapping initiation sites of DNA replication in vivo using polymerase chain reaction amplification of nascent strand segments.
    Nucleic Acids Res. 1989 Oct 11;17(19):7693-705 PMID: 2552413
  11. Thermal energy suppresses mutational defects in DNA unwinding at a yeast replication origin.
    Proc Natl Acad Sci U S A. 1990 Apr;87(7):2486-90 PMID: 2181439
  12. Multiple replication origins are used during Drosophila chorion gene amplification.
    J Cell Biol. 1990 Apr;110(4):903-14 PMID: 2157721
  13. Start sites of bidirectional DNA synthesis at the human lamin B2 origin.
    Science. 2000 Mar 17;287(5460):2023-6 PMID: 10720330
  14. Major DNA replication initiation sites in the c-myc locus in human cells.
    J Cell Biochem. 2000 Jun 6;78(3):442-57 PMID: 10861842
  15. Mapping an origin of DNA replication at a single-copy locus in exponentially proliferating mammalian cells.
    Mol Cell Biol. 1990 Sep;10(9):4685-9 PMID: 2388621
  16. An initiation zone of chromosomal DNA replication located upstream of the c-myc gene in proliferating HeLa cells.
    Mol Cell Biol. 1990 Sep;10(9):4899-904 PMID: 2201906
  17. Emetine allows identification of origins of mammalian DNA replication by imbalanced DNA synthesis, not through conservative nucleosome segregation.
    EMBO J. 1991 Dec;10(13):4351-60 PMID: 1721870
  18. Guide to identification of origins of DNA replication in eukaryotic cell chromosomes.
    Crit Rev Biochem Mol Biol. 1992;27(6):445-72 PMID: 1473351
  19. A novel procedure for quantitative polymerase chain reaction by coamplification of competitive templates.
    Gene. 1992 Dec 15;122(2):313-20 PMID: 1487146
  20. Immunoglobulin heavy chain enhancer is located near or in an initiation zone of chromosomal DNA replication.
    Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3695-9 PMID: 8475117
  21. DNA replication of histone gene repeats in Drosophila melanogaster tissue culture cells: multiple initiation sites and replication pause sites.
    Mol Cell Biol. 1993 Jul;13(7):4098-106 PMID: 8321216
  22. Analysis of a replication initiation sequence from the adenosine deaminase region of the mouse genome.
    Mol Cell Biol. 1993 Oct;13(10):5931-42 PMID: 8413198
  23. Initiation and termination of DNA replication in human rRNA genes.
    Mol Cell Biol. 1993 Oct;13(10):6600-13 PMID: 8413256
  24. A DNA unwinding element and an ARS consensus comprise a replication origin within a yeast chromosome.
    EMBO J. 1993 Dec;12(12):4521-31 PMID: 8223462
  25. Replication structure of the human beta-globin gene domain.
    Nature. 1993 Dec 9;366(6455):588-90 PMID: 8255298
  26. Fine mapping of a replication origin of human DNA.
    Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7119-23 PMID: 8041756
  27. Mapping an initiation region of DNA replication at a single-copy chromosomal locus in Drosophila melanogaster cells by two-dimensional gel methods and PCR-mediated nascent-strand analysis: multiple replication origins in a broad zone.
    Mol Cell Biol. 1994 Nov;14(11):7394-403 PMID: 7935453
  28. Site-specific initiation of DNA replication in Xenopus egg extract requires nuclear structure.
    Mol Cell Biol. 1995 Jun;15(6):2942-54 PMID: 7760792
  29. The Chinese hamster dihydrofolate reductase origin consists of multiple potential nascent-strand start sites.
    Mol Cell Biol. 1995 Jun;15(6):3023-31 PMID: 7760799
  30. Participation of the human beta-globin locus control region in initiation of DNA replication.
    Science. 1995 Nov 3;270(5237):815-9 PMID: 7481774
  31. Mathematical considerations of competitive polymerase chain reaction.
    J Immunol Methods. 1995 Dec 1;187(2):201-11 PMID: 7499879
  32. DNA replication initiates non-randomly at multiple sites near the c-myc gene in HeLa cells.
    Nucleic Acids Res. 1996 May 15;24(10):1887-94 PMID: 8657570
  33. Utilization of the same DNA replication origin by human cells of different derivation.
    Nucleic Acids Res. 1996 Sep 1;24(17):3289-94 PMID: 8811079
  34. High-resolution mapping of the origin of DNA replication in the hamster dihydrofolate reductase gene domain by competitive PCR.
    Mol Cell Biol. 1996 Oct;16(10):5358-64 PMID: 8816447
  35. Chromosome architecture can dictate site-specific initiation of DNA replication in Xenopus egg extracts.
    J Cell Biol. 1996 Dec;135(5):1207-18 PMID: 8947545
  36. Differential DNA replication origin activities in human normal skin fibroblast and HeLa cell lines.
    J Mol Biol. 1997 Oct 31;273(3):509-18 PMID: 9356241
  37. Initiation of DNA replication at CpG islands in mammalian chromosomes.
    EMBO J. 1998 Apr 15;17(8):2426-35 PMID: 9545253
  38. Identification of primary initiation sites for DNA replication in the hamster dihydrofolate reductase gene initiation zone.
    Mol Cell Biol. 1998 Jun;18(6):3266-77 PMID: 9584167
  39. An initiation zone of chromosomal DNA replication at the chicken lysozyme gene locus.
    J Biol Chem. 1998 Jul 17;273(29):18300-7 PMID: 9660795
  40. Genetic dissection of a mammalian replicator in the human beta-globin locus.
    Science. 1998 Aug 14;281(5379):1005-9 PMID: 9703500
  41. Regulation of mammalian replication origin usage in Xenopus egg extract.
    J Cell Sci. 1998 Oct;111 ( Pt 19):2989-98 PMID: 9730990
  42. Multiple initiations in the c-myc replication origin independent of chromosomal location.
    DNA Cell Biol. 1998 Oct;17(10):885-96 PMID: 9809750
  43. Identification of initiation sites for DNA replication in the human dnmt1 (DNA-methyltransferase) locus.
    J Biol Chem. 1999 Apr 2;274(14):9335-41 PMID: 10092611
  44. Activity of the c-myc replicator at an ectopic chromosomal location.
    Mol Cell Biol. 1999 Aug;19(8):5685-95 PMID: 10409757
  45. ORC binding, gene amplification, and the nature of metazoan replication origins.
    Genes Dev. 1999 Oct 15;13(20):2619-23 PMID: 10541547
  46. Drosophila ORC specifically binds to ACE3, an origin of DNA replication control element.
    Genes Dev. 1999 Oct 15;13(20):2639-49 PMID: 10541550
  47. An origin of bidirectional DNA replication is located within a CpG island at the 3" end of the chicken lysozyme gene.
    Nucleic Acids Res. 1999 Aug 1;27(15):3009-17 PMID: 10454594
  48. DNA methylation at mammalian replication origins.
    J Biol Chem. 1999 Sep 3;274(36):25792-800 PMID: 10464318
  49. Long-distance control of origin choice and replication timing in the human beta-globin locus are independent of the locus control region.
    Mol Cell Biol. 2000 Aug;20(15):5581-91 PMID: 10891496
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/29.3.809
Published
2001-02-01
Pages
809-17
Language
English
Region
England
NLM ID
0411011
PMCID
PMC30394
Subset
IM
Grants
NIGMS NIH HHS · R01 GM053819 · United States
NIGMS NIH HHS · GM53819 · United States
External Links
PubMed source DOI: 10.1093/nar/29.3.809 PMC Full Text View journal details
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