Home LiteratureArticle Details
PMID: 230506 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Single-strand binding protein enhances fidelity of DNA synthesis in vitro.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 76 ·No. 12 ·1979-12-00 ·Pages 6331-5

Kunkel TA, Meyer RR, Loeb LA

Abstract

The effect of Escherichia coli single-strand binding protein on the accuracy of in vitro DNA synthesis has been determined by using two independent methods. By using the synthetic polynucleotide poly[d(A-T)] and measuring dGTP misincorporation or by using phi X174 DNA and measuring nucleotide substitutions, we found that binding protein increases the fidelity of DNA synthesis by as much as 10-fold. This increase is observed with DNA polymerases of divergent sources and is progressive with increasing concentration of binding protein. The increased accuracy observed with DNA polymerases lacking a 3' leads to 5' exonuclease points to a mechanism other than augmented proofreading. In accord with the properties of single-strand binding proteins, it is suggested that increased fidelity is a result of enhanced base selection by the DNA polymerase, resulting from increased rigidity of the template due to its interaction with binding protein.

MeSH Terms
Base Sequence DNA Helicases/metabolism DNA Polymerase III/metabolism DNA Replication DNA, Bacterial/metabolism DNA, Single-Stranded/metabolism DNA-Directed DNA Polymerase/metabolism Escherichia coli/genetics Nucleic Acid Conformation Substrate Specificity Templates, Genetic
Chemicals
DNA, Bacterial DNA, Single-Stranded DNA Polymerase III DNA-Directed DNA Polymerase DNA Helicases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kunkel T A
Meyer R R
Loeb L A
References (41)
41 references, click to expand
  1. Control of mutation frequency by bacteriophage T4 DNA polymerase. II. Accuracy of nucleotide selection by the L88 mutator, CB120 antimutator, and wild type phage T4 DNA polymerases.
    J Biol Chem. 1976 Sep 10;251(17):5225-32 PMID: 956183
  2. Deoxyribonucleic acid polymerase III of Escherichia coli. Purification and properties.
    J Biol Chem. 1975 Jan 25;250(2):461-9 PMID: 1089643
  3. Purification and physical characterization of nucleic acid helix-unwinding proteins from calf thymus.
    J Biol Chem. 1976 Apr 10;251(7):2124-32 PMID: 1270425
  4. The effect of nitrosoguanidine upon DNA synthesis in vitro.
    Mol Gen Genet. 1976 May 7;145(2):113-7 PMID: 934048
  5. T4 bacteriophage gene 32: a structural protein in the replication and recombination of DNA.
    Nature. 1970 Sep 26;227(5265):1313-8 PMID: 5455134
  6. Probing DNA replication mechanisms with the T4 bacteriophage in vitro system.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:469-87 PMID: 225114
  7. On the fidelity of DNA replication.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 2:921-7 PMID: 90579
  8. A new approach to DNA polymerase kinetics.
    J Mol Biol. 1979 Mar 25;129(1):93-112 PMID: 221661
  9. On the fidelity of DNA replication. Effect of divalent metal ion activators and deoxyrionucleoside triphosphate pools on in vitro mutagenesis.
    J Biol Chem. 1979 Jul 10;254(13):5718-25 PMID: 376517
  10. On the fidelity of DNA replication. Studies with human placenta DNA polymerases.
    J Biol Chem. 1979 Jun 25;254(12):5229-37 PMID: 447644
  11. DNA polymerase III of Escherichia coli. Purification and identification of subunits.
    J Biol Chem. 1979 Mar 10;254(5):1748-53 PMID: 368075
  12. Mutagenesis in vitro by DNA polymerase from an RNA tumour virus.
    Nature. 1979 Apr 26;278(5707):857-9 PMID: 220540
  13. Mutagenesis during in vitro DNA synthesis.
    Proc Natl Acad Sci U S A. 1978 Apr;75(4):1924-8 PMID: 347450
  14. On the fidelity of DNA replication. Accuracy of Escherichia coli DNA polymerase I.
    J Biol Chem. 1979 Jan 10;254(1):101-6 PMID: 363714
  15. Effect of metal cations on misincorporation by E. coli DNA polymerases.
    Biochem Biophys Res Commun. 1977 Aug 8;77(3):854-60 PMID: 332171
  16. Eukaryotic DNA polymerases.
    Annu Rev Biochem. 1977;46:25-47 PMID: 332062
  17. A deoxyribonucleic acid unwinding protein isolated from regenerating rat liver. Physical and functional properties.
    J Biol Chem. 1978 Mar 10;253(5):1660-6 PMID: 203598
  18. DNA polymerase III holoenzyme of Escherichia coli. Purification and resolution into subunits.
    J Biol Chem. 1977 Sep 25;252(18):6478-84 PMID: 330531
  19. An Escherichia coli mutant defective in single-strand binding protein is defective in DNA replication.
    Proc Natl Acad Sci U S A. 1979 Apr;76(4):1702-5 PMID: 221903
  20. Enzymatic conversion of single-stranded phiX174 and G4 circles to duplex forms: discontinuous replication.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:289-93 PMID: 225101
  21. Mutator and antimutator phenotypes of suppressed amber mutants in genes 32, 41, 44, 45, and 62 in bacteriophage T4.
    J Virol. 1978 Jan;25(1):73-7 PMID: 621787
  22. Multiple interactions of a DNA-binding protein in vivo. I. Gene 32 mutations of phage T4 inactivate different steps in DNA replication and recombination.
    J Mol Biol. 1977 May 15;112(2):279-94 PMID: 889598
  23. DNA unwinding component of the nonhistone chromatin proteins.
    Proc Natl Acad Sci U S A. 1976 Dec;73(12):4364-8 PMID: 1069988
  24. Infidelity of DNA synthesis in vitro: screening for potential metal mutagens or carcinogens.
    Science. 1976 Dec 24;194(4272):1434-6 PMID: 1006310
  25. Conformation of deoxynucleoside triphosphate substrates on DNA polymerase I from Escherichia coli as determined by nuclear magnetic relaxation.
    J Biol Chem. 1975 Dec 10;250(23):8913-20 PMID: 1104609
  26. Novikoff hepatoma deoxyribonucleic acid polymerase. Purification and properties of a homogeneous beta polymerase.
    Biochemistry. 1976 Jul 13;15(14):3114-21 PMID: 182203
  27. Purification and properties of the Escherichia coli deoxyribonucleic acid-unwinding protein. Effects on deoxyribonucleic acid synthesis in vitro.
    J Biol Chem. 1974 Oct 10;249(19):6090-8 PMID: 4608147
  28. Deoxyribonucleic acid synthesis in cell-free extracts. IV. Purification and catalytic properties of deoxyribonucleic acid polymerase III.
    J Biol Chem. 1972 Sep 10;247(17):5369-75 PMID: 4560196
  29. The infidelity of avian myeloblastosis virus deoxyribonucleic acid polymerase in polynucleotide replication.
    J Biol Chem. 1974 Jul 10;249(13):4086-93 PMID: 4137275
  30. Comparative rates of spontaneous mutation.
    Nature. 1969 Mar 22;221(5186):1132 PMID: 4378427
  31. The isolation and properties of a DNA-unwinding protein from Ustilago maydis.
    J Mol Biol. 1975 Mar 25;93(1):63-77 PMID: 1152048
  32. On the fidelity of DNA replication. Lack of exodeoxyribonuclease activity and error-correcting function in avian myeloblastosis virus DNA polymerase.
    J Biol Chem. 1976 Feb 25;251(4):982-6 PMID: 55415
  33. Escherichia coli deoxyribonucleic acid polymerase I, a zinc metalloenzyme. Nuclear quadrupolar relaxation studies of the role of bound zinc.
    J Biol Chem. 1973 Sep 10;248(17):5987-93 PMID: 4580048
  34. A DNA-unwinding protein isolated from Escherichia coli: its interaction with DNA and with DNA polymerases.
    Proc Natl Acad Sci U S A. 1972 Dec;69(12):3537-41 PMID: 4566449
  35. Stimulation of mutation in phage T 4 by lesions in gene 32 and by thymidine imbalance.
    Mutat Res. 1972 Oct;16(2):113-9 PMID: 4561494
  36. Avian myeloblastosis virus DNA polymerase. Kinetic studies on the incorporation of noncomplementary nucleotides.
    J Biol Chem. 1975 Nov 10;250(21):8404-8 PMID: 172498
  37. Studies on a mammalian cell protein (P8) with affinity for DNA in vitro.
    J Mol Biol. 1973 Feb 19;73(3):307-16 PMID: 4686197
  38. An in vitro transversion by a mutationally altered T4-induced DNA polymerase.
    J Mol Biol. 1968 Sep 28;36(3):321-33 PMID: 4939629
  39. Analysis of DNA polymerases II and 3 in mutants of Escherichia coli thermosensitive for DNA synthesis.
    Proc Natl Acad Sci U S A. 1971 Dec;68(12):3150-3 PMID: 4943556
  40. The deoxyribonucleic acid unwinding protein of Escherichia coli. Properties and functions in replication.
    J Biol Chem. 1975 Mar 25;250(6):1972-80 PMID: 1090613
  41. Mutator mutations in bacteriophage T4 gene 32 (DNA unwinding protein).
    J Virol. 1976 Aug;19(2):490-4 PMID: 957480
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.76.12.6331
Published
1979-12-00
Pages
6331-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC411858
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.76.12.6331 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: product@genelibs.com

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