Skip to main content
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
. 1994 Jul 19;91(15):7119–7123. doi: 10.1073/pnas.91.15.7119

Fine mapping of a replication origin of human DNA.

M Giacca 1, L Zentilin 1, P Norio 1, S Diviacco 1, D Dimitrova 1, G Contreas 1, G Biamonti 1, G Perini 1, F Weighardt 1, S Riva 1, et al.
PMCID: PMC44350  PMID: 8041756

Abstract

A highly sensitive procedure was developed for the identification of the origin of bidirectional DNA synthesis in single-copy replicons of mammalian cells. The method, which does not require cell synchronization or permeabilization, entails the absolute quantification, by a competitive PCR procedure in newly synthesized DNA samples, of the abundance of neighboring DNA fragments distributed along a given genomic region. This procedure was utilized for mapping the start site of DNA replication in a 13.7-kb region of human chromosome 19 coding for lamin B2, which is replicated immediately after the onset of S phase in HL-60 cells. Within this region, DNA replication initiates in a 474-bp area corresponding to the 3' noncoding end of the lamin B2 gene and the nontranscribed spacer between this gene and the 5' end of another highly transcribed one. This localization was obtained both in aphidicolin-synchronized and in exponentially growing HL-60 cells.

Full text

PDF
7123

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anson D. S., Choo K. H., Rees D. J., Giannelli F., Gould K., Huddleston J. A., Brownlee G. G. The gene structure of human anti-haemophilic factor IX. EMBO J. 1984 May;3(5):1053–1060. doi: 10.1002/j.1460-2075.1984.tb01926.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barker K. A., Orkin S. H., Newburger P. E. Expression of the X-CGD gene during induced differentiation of myeloid leukemia cell line HL-60. Mol Cell Biol. 1988 Jul;8(7):2804–2810. doi: 10.1128/mcb.8.7.2804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benbow R. M., Zhao J., Larson D. D. On the nature of origins of DNA replication in eukaryotes. Bioessays. 1992 Oct;14(10):661–670. doi: 10.1002/bies.950141004. [DOI] [PubMed] [Google Scholar]
  4. Biamonti G., Giacca M., Perini G., Contreas G., Zentilin L., Weighardt F., Guerra M., Della Valle G., Saccone S., Riva S. The gene for a novel human lamin maps at a highly transcribed locus of chromosome 19 which replicates at the onset of S-phase. Mol Cell Biol. 1992 Aug;12(8):3499–3506. doi: 10.1128/mcb.12.8.3499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Biamonti G., Perini G., Weighardt F., Riva S., Giacca M., Norio P., Zentilin L., Diviacco S., Dimitrova D., Falaschi A. A human DNA replication origin: localization and transcriptional characterization. Chromosoma. 1992;102(1 Suppl):S24–S31. doi: 10.1007/BF02451782. [DOI] [PubMed] [Google Scholar]
  6. Burhans W. C., Vassilev L. T., Caddle M. S., Heintz N. H., DePamphilis M. L. Identification of an origin of bidirectional DNA replication in mammalian chromosomes. Cell. 1990 Sep 7;62(5):955–965. doi: 10.1016/0092-8674(90)90270-o. [DOI] [PubMed] [Google Scholar]
  7. Caddle M. S., Calos M. P. Analysis of the autonomous replication behavior in human cells of the dihydrofolate reductase putative chromosomal origin of replication. Nucleic Acids Res. 1992 Nov 25;20(22):5971–5978. doi: 10.1093/nar/20.22.5971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Contreas G., Giacca M., Falaschi A. Purification of BrdUrd-substituted DNA by immunoaffinity chromatography with anti-BrdUrd antibodies. Biotechniques. 1992 Jun;12(6):824–826. [PubMed] [Google Scholar]
  9. Dalla-Favera R., Wong-Staal F., Gallo R. C. Onc gene amplification in promyelocytic leukaemia cell line HL-60 and primary leukaemic cells of the same patient. Nature. 1982 Sep 2;299(5878):61–63. doi: 10.1038/299061a0. [DOI] [PubMed] [Google Scholar]
  10. DePamphilis M. L. Transcriptional elements as components of eukaryotic origins of DNA replication. Cell. 1988 Mar 11;52(5):635–638. doi: 10.1016/0092-8674(88)90398-4. [DOI] [PubMed] [Google Scholar]
  11. Diviacco S., Norio P., Zentilin L., Menzo S., Clementi M., Biamonti G., Riva S., Falaschi A., Giacca M. A novel procedure for quantitative polymerase chain reaction by coamplification of competitive templates. Gene. 1992 Dec 15;122(2):313–320. doi: 10.1016/0378-1119(92)90220-j. [DOI] [PubMed] [Google Scholar]
  12. Falaschi A., Giacca M., Zentilin L., Norio P., Diviacco S., Dimitrova D., Kumar S., Tuteja R., Biamonti G., Perini G. Searching for replication origins in mammalian DNA. Gene. 1993 Dec 15;135(1-2):125–135. doi: 10.1016/0378-1119(93)90057-a. [DOI] [PubMed] [Google Scholar]
  13. Gilbert D., Cohen S. N. Autonomous replication in mouse cells: a correction. Cell. 1989 Jan 27;56(2):143–144. doi: 10.1016/0092-8674(89)90885-4. [DOI] [PubMed] [Google Scholar]
  14. Handeli S., Klar A., Meuth M., Cedar H. Mapping replication units in animal cells. Cell. 1989 Jun 16;57(6):909–920. doi: 10.1016/0092-8674(89)90329-2. [DOI] [PubMed] [Google Scholar]
  15. Heintz N. H. Transcription factors and the control of DNA replication. Curr Opin Cell Biol. 1992 Jun;4(3):459–467. doi: 10.1016/0955-0674(92)90012-2. [DOI] [PubMed] [Google Scholar]
  16. Heinzel S. S., Krysan P. J., Tran C. T., Calos M. P. Autonomous DNA replication in human cells is affected by the size and the source of the DNA. Mol Cell Biol. 1991 Apr;11(4):2263–2272. doi: 10.1128/mcb.11.4.2263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Huberman J. A., Riggs A. D. On the mechanism of DNA replication in mammalian chromosomes. J Mol Biol. 1968 Mar 14;32(2):327–341. doi: 10.1016/0022-2836(68)90013-2. [DOI] [PubMed] [Google Scholar]
  18. Kitsberg D., Selig S., Keshet I., Cedar H. Replication structure of the human beta-globin gene domain. Nature. 1993 Dec 9;366(6455):588–590. doi: 10.1038/366588a0. [DOI] [PubMed] [Google Scholar]
  19. Kowalski D., Eddy M. J. The DNA unwinding element: a novel, cis-acting component that facilitates opening of the Escherichia coli replication origin. EMBO J. 1989 Dec 20;8(13):4335–4344. doi: 10.1002/j.1460-2075.1989.tb08620.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lüscher B., Eisenman R. N. New light on Myc and Myb. Part I. Myc. Genes Dev. 1990 Dec;4(12A):2025–2035. doi: 10.1101/gad.4.12a.2025. [DOI] [PubMed] [Google Scholar]
  21. Prendergast G. C., Ziff E. B. A new bind for Myc. Trends Genet. 1992 Mar;8(3):91–96. doi: 10.1016/0168-9525(92)90196-b. [DOI] [PubMed] [Google Scholar]
  22. Tribioli C., Biamonti G., Giacca M., Colonna M., Riva S., Falaschi A. Characterization of human DNA sequences synthesized at the onset of S-phase. Nucleic Acids Res. 1987 Dec 23;15(24):10211–10232. doi: 10.1093/nar/15.24.10211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tóth E. C., Marusic L., Ochem A., Patthy A., Pongor S., Giacca M., Falaschi A. Interactions of USF and Ku antigen with a human DNA region containing a replication origin. Nucleic Acids Res. 1993 Jul 11;21(14):3257–3263. doi: 10.1093/nar/21.14.3257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Umek R. M., Kowalski D. The ease of DNA unwinding as a determinant of initiation at yeast replication origins. Cell. 1988 Feb 26;52(4):559–567. doi: 10.1016/0092-8674(88)90469-2. [DOI] [PubMed] [Google Scholar]
  25. Vassilev L. T., Burhans W. C., DePamphilis M. L. Mapping an origin of DNA replication at a single-copy locus in exponentially proliferating mammalian cells. Mol Cell Biol. 1990 Sep;10(9):4685–4689. doi: 10.1128/mcb.10.9.4685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Vassilev L., Johnson E. M. 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):4899–4904. doi: 10.1128/mcb.10.9.4899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Vaughn J. P., Dijkwel P. A., Hamlin J. L. Replication initiates in a broad zone in the amplified CHO dihydrofolate reductase domain. Cell. 1990 Jun 15;61(6):1075–1087. doi: 10.1016/0092-8674(90)90071-l. [DOI] [PubMed] [Google Scholar]
  28. Zhu J., Brun C., Kurooka H., Yanagida M., Huberman J. A. Identification and characterization of a complex chromosomal replication origin in Schizosaccharomyces pombe. Chromosoma. 1992;102(1 Suppl):S7–16. doi: 10.1007/BF02451780. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES