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PMID: 8371975 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

A general protocol for evaluating the specific effects of DNA replication inhibitors.

Nucleic acids research ·Vol. 21 ·No. 17 ·1993-08-25 ·Pages 3997-4004

Levenson V, Hamlin JL

Abstract

Inhibitors of DNA replication in mammalian cells are of great interest because of their potential use in chemotherapy and in cell synchronizing protocols in the laboratory. We have used a combination of isotopic labelling protocols and a two-dimensional gel replicon mapping procedure to determine the specific effects of five different replication inhibitors in cultured cells. Utilizing this protocol, we show that hydroxyurea, aphidicolin, and cytosine arabinoside, three known chain elongation inhibitors, are rather ineffective at preventing fork progression even at relatively high concentrations. In contrast, two related compounds that have been suggested to be G1/S inhibitors (mimosine and ciclopyrox olamine [CPX]) actually appear to inhibit initiation at origins. One of these agents (CPX) appears also to inhibit replication in yeast, opening the possibility that the gene encoding the target (initiator?) protein can first be identified in yeast by genetic approaches and can then be used to isolate the mammalian homologue.

MeSH Terms
Animals Antifungal Agents/pharmacology Aphidicolin/pharmacology CHO Cells Cell Cycle/drug effects Ciclopirox Cricetinae Cytarabine/pharmacology DNA Replication/drug effects Electrophoresis, Gel, Two-Dimensional Flow Cytometry Genetic Techniques Hydroxyurea/pharmacology Mimosine/pharmacology Pyridones/pharmacology Replicon/drug effects Tetrahydrofolate Dehydrogenase/genetics
Chemicals
Antifungal Agents Pyridones Cytarabine Ciclopirox Aphidicolin Mimosine Tetrahydrofolate Dehydrogenase Hydroxyurea
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Levenson V
University of Virginia School of Medicine, Department of Biochemistry, Charlottesville 22908.
Hamlin J L
References (29)
29 references, click to expand
  1. The mechanism of action of inhibitors of DNA synthesis.
    Annu Rev Biochem. 1977;46:641-68 PMID: 143235
  2. S phase synchrony in monolayer CHO cultures.
    Exp Cell Res. 1976 Jul;100(2):265-75 PMID: 939252
  3. 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
  4. 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
  5. In vivo effects of cytosine arabinoside on deoxyribonucleic acid replication in Chinese hamster ovary cells. 1. Resolution of differential effects on mitochondrial and nuclear deoxyribonucleic acid synthesis.
    Biochemistry. 1983 Jul 19;22(15):3552-7 PMID: 6615783
  6. Replication timing of genes and middle repetitive sequences.
    Science. 1984 May 18;224(4650):686-92 PMID: 6719109
  7. Organization of the higher-order chromatin loop: specific DNA attachment sites on nuclear scaffold.
    Cell. 1984 Nov;39(1):223-32 PMID: 6091913
  8. Cloning of nascent monkey DNA synthesized early in the cell cycle.
    Mol Cell Biol. 1985 Apr;5(4):721-7 PMID: 3990692
  9. Ciclopirox olamine 1% cream. A preliminary review of its antimicrobial activity and therapeutic use.
    Drugs. 1985 Apr;29(4):330-41 PMID: 3158508
  10. The localization of replication origins on ARS plasmids in S. cerevisiae.
    Cell. 1987 Nov 6;51(3):463-71 PMID: 2822257
  11. Temporal order of gene replication in Chinese hamster ovary cells.
    Mol Cell Biol. 1989 Jul;9(7):2881-9 PMID: 2476659
  12. A reversible arrest point in the late G1 phase of the mammalian cell cycle.
    Exp Cell Res. 1990 Feb;186(2):332-9 PMID: 2153561
  13. Replication forks are associated with the nuclear matrix.
    Nucleic Acids Res. 1990 Apr 25;18(8):1965-9 PMID: 2336385
  14. Replication initiates in a broad zone in the amplified CHO dihydrofolate reductase domain.
    Cell. 1990 Jun 15;61(6):1075-87 PMID: 2350784
  15. Identification of an origin of bidirectional DNA replication in mammalian chromosomes.
    Cell. 1990 Sep 7;62(5):955-65 PMID: 2393905
  16. A new class of reversible cell cycle inhibitors.
    Cytometry. 1991;12(1):26-32 PMID: 1900227
  17. 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
  18. Eukaryotic DNA polymerases.
    Annu Rev Biochem. 1991;60:513-52 PMID: 1883203
  19. 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
  20. The plant amino acid mimosine may inhibit initiation at origins of replication in Chinese hamster cells.
    Mol Cell Biol. 1992 Oct;12(10):4375-83 PMID: 1406627
  21. Antimetabolites.
    Curr Opin Oncol. 1992 Dec;4(6):1089-98 PMID: 1457523
  22. Studies on unbalanced growth in tissue culture. I. Induction and consequences of thymidine deficiency.
    Cancer Res. 1960 Dec;20:1584-91 PMID: 13744520
  23. SOME MECHANISMS INVOLVED IN SELECTIVE CHEMOTHERAPY.
    Ann N Y Acad Sci. 1965 Mar 12;123:19-41 PMID: 14329203
  24. On the mechanism of DNA replication in mammalian chromosomes.
    J Mol Biol. 1968 Mar 14;32(2):327-41 PMID: 5689363
  25. DNA replication in SV40 infected cells. I. Analysis of replicating SV40 DNA.
    J Mol Biol. 1970 Jun 14;50(2):549-68 PMID: 4320050
  26. Effects of hydroxyurea and 1-beta-D-arabinofuranosyl-cytosine on deoxyribonucleotide pools in mouse embryo cells.
    Eur J Biochem. 1971 Mar 1;19(1):81-9 PMID: 5573224
  27. Escherichia coli mutants temperature-sensitive for DNA synthesis.
    Mol Gen Genet. 1971;113(3):273-84 PMID: 4946856
  28. Preparation of large quantities of synchronized mammalian cells in late G1 in the pre-DNA replicative phase of the cell cycle.
    Exp Cell Res. 1972 Dec;75(2):460-4 PMID: 4674697
  29. Synchronization of HeLa cell cultures by inhibition of DNA polymerase alpha with aphidicolin.
    Nucleic Acids Res. 1980 Jan 25;8(2):377-87 PMID: 6775308
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/21.17.3997
Published
1993-08-25
Pages
3997-4004
Language
English
Region
England
NLM ID
0411011
PMCID
PMC309986
Subset
IM
Grants
NCI NIH HHS · R01CA52559 · United States
NIGMS NIH HHS · R01GM26108 · United States
External Links
PubMed source DOI: 10.1093/nar/21.17.3997 PMC Full Text View journal details
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