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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
. 1973 Jun;70(6):1846–1850. doi: 10.1073/pnas.70.6.1846

A DNA-Binding Protein Induced by Bacteriophage T7

Roberta C Reuben *,*, Malcolm L Gefter
PMCID: PMC433610  PMID: 4578445

Abstract

A DNA-binding protein has been purified from Escherichia coli infected with bacteriophage T7 by DNA-cellulose chromatography. The protein is absent in uninfected cells. The purified protein has a molecular weight of 31,000 and binds strongly and preferentially to single-stranded DNA. In vitro studies show that this protein can stimulate the rate of polymerization catalyzed by the T7-induced DNA polymerase 10-15 times under conditions where the polymerase is unable to effectively use a single-stranded template. The degree of stimulation is dependent upon the ratio of binding protein to DNA template and is independent of polymerase concentration.

The observed stimulation is specific for the T7 DNA polymerase in that addition of the protein to reactions catalyzed by E. coli DNA polymerases I, II, or III or T4 DNA polymerase is without effect.

Keywords: DNA cellulose, T7 DNA polymerase, stimulation of synthesis

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Selected References

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

  1. Alberts B. M., Amodio F. J., Jenkins M., Gutmann E. D., Ferris F. L. Studies with DNA-cellulose chromatography. I. DNA-binding proteins from Escherichia coli. Cold Spring Harb Symp Quant Biol. 1968;33:289–305. doi: 10.1101/sqb.1968.033.01.033. [DOI] [PubMed] [Google Scholar]
  2. Alberts B. M., Frey L. T4 bacteriophage gene 32: a structural protein in the replication and recombination of DNA. Nature. 1970 Sep 26;227(5265):1313–1318. doi: 10.1038/2271313a0. [DOI] [PubMed] [Google Scholar]
  3. Grippo P., Richardson C. C. Deoxyribonucleic acid polymerase of bacteriophage T7. J Biol Chem. 1971 Nov 25;246(22):6867–6873. [PubMed] [Google Scholar]
  4. Huberman J. A., Kornberg A., Alberts B. M. Stimulation of T4 bacteriophage DNA polymerase by the protein product of T4 gene 32. J Mol Biol. 1971 Nov 28;62(1):39–52. doi: 10.1016/0022-2836(71)90129-x. [DOI] [PubMed] [Google Scholar]
  5. Kornberg T., Gefter M. L. 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):5369–5375. [PubMed] [Google Scholar]
  6. Kornberg T., Gefter M. L. Purification and DNA synthesis in cell-free extracts: properties of DNA polymerase II. Proc Natl Acad Sci U S A. 1971 Apr;68(4):761–764. doi: 10.1073/pnas.68.4.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Oey J. L., Strätling W., Knippers R. A DNA polymerase induced by bacteriophage T7. Eur J Biochem. 1971 Dec 10;23(3):497–504. doi: 10.1111/j.1432-1033.1971.tb01646.x. [DOI] [PubMed] [Google Scholar]
  8. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  9. Sigal N., Delius H., Kornberg T., Gefter M. L., Alberts B. 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):3537–3541. doi: 10.1073/pnas.69.12.3537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Sinha N. K., Snustad D. P. DNA synthesis in bacteriophage T4-infected Escherichia coli: evidence supporting a stoichiometric role for gene 32-product. J Mol Biol. 1971 Nov 28;62(1):267–271. doi: 10.1016/0022-2836(71)90145-8. [DOI] [PubMed] [Google Scholar]
  11. Studier F. W. Bacteriophage T7. Science. 1972 Apr 28;176(4033):367–376. doi: 10.1126/science.176.4033.367. [DOI] [PubMed] [Google Scholar]
  12. Wechsler J. A., Gross J. D. Escherichia coli mutants temperature-sensitive for DNA synthesis. Mol Gen Genet. 1971;113(3):273–284. doi: 10.1007/BF00339547. [DOI] [PubMed] [Google Scholar]

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