Home LiteratureArticle Details
PMID: 8816447 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

High-resolution mapping of the origin of DNA replication in the hamster dihydrofolate reductase gene domain by competitive PCR.

Molecular and cellular biology ·Vol. 16 ·No. 10 ·1996-10-00 ·Pages 5358-64

Pelizon C, Diviacco S, Falaschi A, Giacca M

Abstract

By the use of a highly sensitive mapping procedure allowing the identification of the start sites of DNA replication in single-copy genomic regions of untreated, exponentially growing cultured cells (M. Giacca, L. Zentilin, P. Norio, S. Diviacco, D. Dimitrova, G. Contreas, G. Biamonti, G. Perini, F. Weighardt, S. Riva, and A. Falaschi, Proc. Natl. Acad. Sci. USA 91:7119-7123, 1994), the pattern of DNA replication of the Chinese hamster dihydrofolate reductase (DHFR) gene domain was investigated. The method entails the purification of short stretches of nascent DNA issuing from DNA replication origin regions and quantification, within this sample, of the abundance of different adjacent segments by competitive PCR. Distribution of marker abundance peaks around the site from which newly synthesized DNA had emanated. The results obtained by analysis of the genomic region downstream of the DHFR single-copy gene in asynchronous cultures of hamster CHO K1 cells are consistent with the presence of a single start site for DNA replication, located approximately 17 kb downstream of the gene. This site is coincident with the one detected by other studies using different techniques in CHO cell lines containing an amplified DHFR gene domain.

MeSH Terms
Animals Base Sequence CHO Cells Chromosome Mapping Cricetinae Cricetulus DNA/analysis DNA Primers DNA Replication Molecular Sequence Data Polymerase Chain Reaction/methods Replication Origin Restriction Mapping Tetrahydrofolate Dehydrogenase/biosynthesis,genetics
Chemicals
DNA Primers DNA Tetrahydrofolate Dehydrogenase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Pelizon C
International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
Diviacco S
Falaschi A
Giacca M
References (43)
43 references, click to expand
  1. Mapping of replication initiation sites in mammalian genomes by two-dimensional gel analysis: stabilization and enrichment of replication intermediates by isolation on the nuclear matrix.
    Mol Cell Biol. 1991 Aug;11(8):3850-9 PMID: 2072896
  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. On the mechanism of DNA replication in mammalian chromosomes.
    J Mol Biol. 1968 Mar 14;32(2):327-41 PMID: 5689363
  4. Isolation and characterisation of a yeast chromosomal replicator.
    Nature. 1979 Nov 1;282(5734):39-43 PMID: 388229
  5. Methotrexate-resistant Chinese hamster ovary cells have amplified a 135-kilobase-pair region that includes the dihydrofolate reductase gene.
    Proc Natl Acad Sci U S A. 1981 Oct;78(10):6043-7 PMID: 6273843
  6. An amplified chromosomal sequence that includes the gene for dihydrofolate reductase initiates replication within specific restriction fragments.
    Proc Natl Acad Sci U S A. 1982 Jul;79(13):4083-7 PMID: 6955792
  7. Fate of exogenous recombinant plasmids introduced into mouse and human cells.
    Nucleic Acids Res. 1985 Aug 12;13(15):5545-61 PMID: 4034393
  8. Isolation of the origin of replication associated with the amplified Chinese hamster dihydrofolate reductase domain.
    Proc Natl Acad Sci U S A. 1986 Oct;83(20):7790-4 PMID: 3094015
  9. Autonomous replication in mouse cells: a correction.
    Cell. 1989 Jan 27;56(2):143-4 PMID: 2912571
  10. High-resolution mapping of replication fork movement through the amplified dihydrofolate reductase domain in CHO cells by in-gel renaturation analysis.
    Mol Cell Biol. 1989 Feb;9(2):523-31 PMID: 2710115
  11. Replication in the amplified dihydrofolate reductase domain in CHO cells may initiate at two distinct sites, one of which is a repetitive sequence element.
    Mol Cell Biol. 1989 Feb;9(2):532-40 PMID: 2710116
  12. Isolation of human sequences that replicate autonomously in human cells.
    Mol Cell Biol. 1989 Mar;9(3):1026-33 PMID: 2542763
  13. Mapping replication units in animal cells.
    Cell. 1989 Jun 16;57(6):909-20 PMID: 2544294
  14. Intramolecular DNA triplexes, bent DNA and DNA unwinding elements in the initiation region of an amplified dihydrofolate reductase replicon.
    J Mol Biol. 1990 Jan 5;211(1):19-33 PMID: 2299670
  15. Replication initiates in a broad zone in the amplified CHO dihydrofolate reductase domain.
    Cell. 1990 Jun 15;61(6):1075-87 PMID: 2350784
  16. 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
  17. 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
  18. The two faces of higher eukaryotic DNA replication origins.
    Cell. 1990 Sep 7;62(5):845-7 PMID: 2203534
  19. Identification of an origin of bidirectional DNA replication in mammalian chromosomes.
    Cell. 1990 Sep 7;62(5):955-65 PMID: 2393905
  20. Replication initiates at multiple locations on an autonomously replicating plasmid in human cells.
    Mol Cell Biol. 1991 Mar;11(3):1464-72 PMID: 1996103
  21. Autonomous DNA replication in human cells is affected by the size and the source of the DNA.
    Mol Cell Biol. 1991 Apr;11(4):2263-72 PMID: 1900922
  22. Purification of BrdUrd-substituted DNA by immunoaffinity chromatography with anti-BrdUrd antibodies.
    Biotechniques. 1992 Jun;12(6):824-6 PMID: 1642886
  23. Initiation of DNA replication in the dihydrofolate reductase locus is confined to the early S period in CHO cells synchronized with the plant amino acid mimosine.
    Mol Cell Biol. 1992 Sep;12(9):3715-22 PMID: 1508178
  24. Competitive PCR.
    Nature. 1992 Oct 8;359(6395):557-8 PMID: 1383831
  25. 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):5971-8 PMID: 1461730
  26. A novel procedure for quantitative polymerase chain reaction by coamplification of competitive templates.
    Gene. 1992 Dec 15;122(2):313-20 PMID: 1487146
  27. Quantitative or semi-quantitative PCR: reality versus myth.
    PCR Methods Appl. 1992 Aug;2(1):1-9 PMID: 1490169
  28. A human DNA replication origin: localization and transcriptional characterization.
    Chromosoma. 1992;102(1 Suppl):S24-31 PMID: 1291239
  29. Localization of a bidirectional DNA replication origin in the native locus and in episomally amplified murine adenosine deaminase loci.
    Mol Cell Biol. 1993 May;13(5):2971-81 PMID: 8474455
  30. Eukaryotic DNA replication: anatomy of an origin.
    Annu Rev Biochem. 1993;62:29-63 PMID: 8352592
  31. 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
  32. Initiation and termination of DNA replication in human rRNA genes.
    Mol Cell Biol. 1993 Oct;13(10):6600-13 PMID: 8413256
  33. Replication structure of the human beta-globin gene domain.
    Nature. 1993 Dec 9;366(6455):588-90 PMID: 8255298
  34. Measuring c-erbB-2 oncogene amplification in fresh and paraffin-embedded tumors by competitive polymerase chain reaction.
    Clin Chem. 1994 Apr;40(4):630-6 PMID: 7511998
  35. 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
  36. A mammalian origin of bidirectional DNA replication within the Chinese hamster RPS14 locus.
    Mol Cell Biol. 1994 Sep;14(9):5628-35 PMID: 8065299
  37. A rapid procedure for the quantitation of low abundance RNAs by competitive reverse transcription-polymerase chain reaction.
    Nucleic Acids Res. 1994 Oct 25;22(21):4547-9 PMID: 7526345
  38. Replication initiation sites are distributed widely in the amplified CHO dihydrofolate reductase domain.
    Nucleic Acids Res. 1994 Nov 25;22(23):4989-96 PMID: 7800491
  39. The quest for a human ori.
    Genetica. 1994;94(2-3):255-66 PMID: 7896144
  40. 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
  41. On the nature of replication origins in higher eukaryotes.
    Curr Opin Genet Dev. 1995 Apr;5(2):153-61 PMID: 7613083
  42. Identification of an origin of bidirectional DNA replication in the ubiquitously expressed mammalian CAD gene.
    Mol Cell Biol. 1995 Aug;15(8):4136-48 PMID: 7623808
  43. In vivo protein-DNA interactions at human DNA replication origin.
    Proc Natl Acad Sci U S A. 1996 Feb 20;93(4):1498-503 PMID: 8643660
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.10.5358
Published
1996-10-00
Pages
5358-64
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231534
Subset
IM
Databases
GENBANK
X94372
External Links
PubMed source DOI: 10.1128/MCB.16.10.5358 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2