Purification and properties of bacteriophage phi X 174 gene D product (original) (raw)

Abstract

Bacteriophage phi X 174 gene D product, a protein required for single-stranded DNA synthesis by the phage, has been purified to near homogeneity. The protein is very abundant; approximately 10(5) monomers are present per infected cell when lysis is delayed. The protein has a monomer molecular weight of 15,200 and is normally a tetramer; however, it can form very large aggregates at high concentrations. Amino acid analysis shows an excess of arginine over lysine and a relatively high number of nonpolar residues. The protein carries a net negative charge at neutral pH. The first eight amino acids of the protein sequence have been determined; they are Ser-Gln-Val-Thr-Glu-Gln-Arg-Val. The carboxy-terminal residue is methionine. The protein has not yet been shown to bind directly to any single-stranded DNA; it does not adsorb to denatured calf thymus DNA-cellulose.

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Selected References

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  1. Alberts B., Frey L., Delius H. Isolation and characterization of gene 5 protein of filamentous bacterial viruses. J Mol Biol. 1972 Jul 14;68(1):139–152. doi: 10.1016/0022-2836(72)90269-0. [DOI] [PubMed] [Google Scholar]
  2. Benbow R. M., Hutchison C. A., Fabricant J. D., Sinsheimer R. L. Genetic Map of Bacteriophage phiX174. J Virol. 1971 May;7(5):549–558. doi: 10.1128/jvi.7.5.549-558.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benbow R. M., Mayol R. F., Picchi J. C., Sinsheimer R. L. Direction of Translation and Size of Bacteriophage phiX174 Cistrons. J Virol. 1972 Jul;10(1):99–114. doi: 10.1128/jvi.10.1.99-114.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burgess A. B., Denhardt D. T. Studies on phiX174 proteins. I. Phage-specific proteins synthesized after infection of Escherichia coli. J Mol Biol. 1969 Sep 28;44(3):377–386. doi: 10.1016/0022-2836(69)90367-2. [DOI] [PubMed] [Google Scholar]
  5. Clements J. B., Sinsheimer R. L. Class of phi chi 174 mutants relatively deficient in synthesis of viral RNA. J Virol. 1974 Dec;14(6):1630–1632. doi: 10.1128/jvi.14.6.1630-1632.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Denhardt D. T., Sinsheimer R. L. The process of infection with bacteriophage phi-X174. IV. Replication of the viral DNA in a synchronized infection. J Mol Biol. 1965 Jul;12(3):647–662. doi: 10.1016/s0022-2836(65)80319-9. [DOI] [PubMed] [Google Scholar]
  7. Dowell C. E., Sinsheimer R. L. The process of infection with bacteriophage phi-X174. IX. Studies on the physiology of three phi-X174 temperature-sensitive mutants. J Mol Biol. 1966 Apr;16(2):374–386. doi: 10.1016/s0022-2836(66)80180-8. [DOI] [PubMed] [Google Scholar]
  8. Edelhoch H. Spectroscopic determination of tryptophan and tyrosine in proteins. Biochemistry. 1967 Jul;6(7):1948–1954. doi: 10.1021/bi00859a010. [DOI] [PubMed] [Google Scholar]
  9. Edman P., Begg G. A protein sequenator. Eur J Biochem. 1967 Mar;1(1):80–91. doi: 10.1007/978-3-662-25813-2_14. [DOI] [PubMed] [Google Scholar]
  10. Gelfand D. H., Hayashi M. Electrophoretic characterization of phiX174-specific proteins. J Mol Biol. 1969 Sep 28;44(3):501–516. doi: 10.1016/0022-2836(69)90376-3. [DOI] [PubMed] [Google Scholar]
  11. Gilbert W., Dressler D. DNA replication: the rolling circle model. Cold Spring Harb Symp Quant Biol. 1968;33:473–484. doi: 10.1101/sqb.1968.033.01.055. [DOI] [PubMed] [Google Scholar]
  12. Godson G. N. Characterization and synthesis of phi X174 proteins in ultraviolet-irradiated and unirradiated cells. J Mol Biol. 1971 May 14;57(3):541–553. doi: 10.1016/0022-2836(71)90108-2. [DOI] [PubMed] [Google Scholar]
  13. Henry T. J., Knippers R. Isolation and function of the gene A initiator of bacteriophage phi-chi 174, a highly specific DNA endonuclease. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1549–1553. doi: 10.1073/pnas.71.4.1549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Horvitz H. R. Polypeptide bound to the host RNA polymerase is specified by T4 control gene 33. Nat New Biol. 1973 Aug 1;244(135):137–140. doi: 10.1038/newbio244137a0. [DOI] [PubMed] [Google Scholar]
  15. Hutchison C. A., 3rd, Sinsheimer R. L. The process of infection with bacteriophage phi-X174. X. Mutations in a phi-X Lysis gene. J Mol Biol. 1966 Jul;18(3):429–447. doi: 10.1016/s0022-2836(66)80035-9. [DOI] [PubMed] [Google Scholar]
  16. Iwaya M., Denhardt D. T. The mechanism of replication of phi X174 single-stranded DNA. II. The role of viral proteins. J Mol Biol. 1971 Apr 28;57(2):159–175. doi: 10.1016/0022-2836(71)90339-1. [DOI] [PubMed] [Google Scholar]
  17. King J., Lenk E. V., Botstein D. Mechanism of head assembly and DNA encapsulation in Salmonella phage P22. II. Morphogenetic pathway. J Mol Biol. 1973 Nov 15;80(4):697–731. doi: 10.1016/0022-2836(73)90205-2. [DOI] [PubMed] [Google Scholar]
  18. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  19. Levine A. J., Sinsheimer R. L. The process of infection with bacteriophage phiX174. XXVII. Synthesis of a viral-specific chloramphenicol-resistant protein in phiX174-infected cells. J Mol Biol. 1969 Feb 14;39(3):655–668. doi: 10.1016/0022-2836(69)90151-x. [DOI] [PubMed] [Google Scholar]
  20. Lindqvist B. H., Sinsheimer R. L. Process of infection with bacteriophage phi-X174. XIV. Studies on macromolecular synthesis during infection with a lysis-defective mutant. J Mol Biol. 1967 Aug 28;28(1):87–94. doi: 10.1016/s0022-2836(67)80079-2. [DOI] [PubMed] [Google Scholar]
  21. Linney E. A., Hayashi M. N., Hayashi M. Gene A of X174. 1. Isolation and identification of its products. Virology. 1972 Nov;50(2):381–387. doi: 10.1016/0042-6822(72)90389-3. [DOI] [PubMed] [Google Scholar]
  22. Mazur B. J., Model P. Regulation of coliphage f1 single-stranded DNA synthesis by a DNA-binding protein. J Mol Biol. 1973 Aug 5;78(2):285–300. doi: 10.1016/0022-2836(73)90117-4. [DOI] [PubMed] [Google Scholar]
  23. Nakashima Y., Dunker A. K., Marvin D. A., Konigsberg W. The amino acid sequence of a DNA binding protein, the gene 5 product of fd filamentous bacteriophage. FEBS Lett. 1974 Apr 1;40(2):290–292. doi: 10.1016/0014-5793(74)80246-2. [DOI] [PubMed] [Google Scholar]
  24. Oey J. L., Knippers R. Properties of the isolated gene 5 protein of bacteriophage fd. J Mol Biol. 1972 Jul 14;68(1):125–138. doi: 10.1016/0022-2836(72)90268-9. [DOI] [PubMed] [Google Scholar]
  25. Platt T., Files J. G., Weber K. Lac repressor. Specific proteolytic destruction of the NH 2 -terminal region and loss of the deoxyribonucleic acid-binding activity. J Biol Chem. 1973 Jan 10;248(1):110–121. [PubMed] [Google Scholar]
  26. Salstrom J. S., Pratt D. Role of coliphage M13 gene 5 in single-stranded DNA production. J Mol Biol. 1971 Nov 14;61(3):489–501. doi: 10.1016/0022-2836(71)90061-1. [DOI] [PubMed] [Google Scholar]
  27. Schekman R., Wickner W., Westergaard O., Brutlag D., Geider K., Bertsch L. L., Kornberg A. Initiation of DNA synthesis: synthesis of phiX174 replicative form requires RNA synthesis resistant to rifampicin. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2691–2695. doi: 10.1073/pnas.69.9.2691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Siden E. J., Hayashi M. Role of the gene beta-product in bacteriophage phi-X174 development. J Mol Biol. 1974 Oct 15;89(1):1–16. doi: 10.1016/0022-2836(74)90159-4. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  31. Weisbeek P. J., Sinsheimer R. L. A DNA-protein complex involved in bacteriophage phi chi 174 particle formation. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3054–3058. doi: 10.1073/pnas.71.8.3054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yamada S., Itano H. Phenanthrenequinone as an analytical reagent for arginine and other monosubstituted guanidines. Biochim Biophys Acta. 1966 Dec 28;130(2):538–540. doi: 10.1016/0304-4165(66)90256-x. [DOI] [PubMed] [Google Scholar]