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. 1991 May 1;88(9):4010–4014. doi: 10.1073/pnas.88.9.4010

Protein-protein interactions with the acidic COOH terminus of the single-stranded DNA-binding protein of the bacteriophage T4.

K B Krassa 1, L S Green 1, L Gold 1
PMCID: PMC51583  PMID: 2023949

Abstract

The single-stranded DNA-binding protein of the bacteriophage T4 is encoded by gene 32. Monoclonal antibodies were raised against intact gene 32 protein (gp32). We mapped the epitopes recognized by 12 of these monoclonal antibodies; the epitopes are all within the COOH-terminal region of gp32. As shown by others, removal of the COOH terminus of gp32 abolishes the ability of the intact protein to bind to many T4 proteins involved in replication, recombination, repair, and late transcription. These results suggest that the COOH terminus of gp32 is a protein-binding domain. The COOH terminus is attached to a DNA-binding domain that includes a zinc finger. We propose a model in which the DNA-binding and protein-binding domains are used in T4 replication, recombination, repair, and late transcription. The COOH terminus of gp32 is very acidic and may form four negatively charged amphipathic alpha-helices, which could fold into a four-helix bundle when associated with other proteins. At least six of the monoclonal anti-gp32 antibodies bind to the COOH terminus of gp32 and to DNA. Similarities between the COOH terminus of gp32 and DNA are explored.

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Selected References

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  1. Alberts B. M., Barry J., Bedinger P., Formosa T., Jongeneel C. V., Kreuzer K. N. Studies on DNA replication in the bacteriophage T4 in vitro system. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 2):655–668. doi: 10.1101/sqb.1983.047.01.077. [DOI] [PubMed] [Google Scholar]
  2. Benjamin D. C., Berzofsky J. A., East I. J., Gurd F. R., Hannum C., Leach S. J., Margoliash E., Michael J. G., Miller A., Prager E. M. The antigenic structure of proteins: a reappraisal. Annu Rev Immunol. 1984;2:67–101. doi: 10.1146/annurev.iy.02.040184.000435. [DOI] [PubMed] [Google Scholar]
  3. Berzofsky J. A. Intrinsic and extrinsic factors in protein antigenic structure. Science. 1985 Sep 6;229(4717):932–940. doi: 10.1126/science.2410982. [DOI] [PubMed] [Google Scholar]
  4. Burke R. L., Alberts B. M., Hosoda J. Proteolytic removal of the COOH terminus of the T4 gene 32 helix-destabilizing protein alters the T4 in vitro replication complex. J Biol Chem. 1980 Dec 10;255(23):11484–11493. [PubMed] [Google Scholar]
  5. Burke R. L., Munn M., Barry J., Alberts B. M. Purification and properties of the bacteriophage T4 gene 61 RNA priming protein. J Biol Chem. 1985 Feb 10;260(3):1711–1722. [PubMed] [Google Scholar]
  6. Carroll R. B., Neet K., Goldthwait D. A. Studies of the self-association of bacteriophage T4 gene 32 protein by equilibrium sedimentation. J Mol Biol. 1975 Jan 25;91(3):275–291. doi: 10.1016/0022-2836(75)90380-0. [DOI] [PubMed] [Google Scholar]
  7. Chase J. W., Williams K. R. Single-stranded DNA binding proteins required for DNA replication. Annu Rev Biochem. 1986;55:103–136. doi: 10.1146/annurev.bi.55.070186.000535. [DOI] [PubMed] [Google Scholar]
  8. Formosa T., Alberts B. M. DNA synthesis dependent on genetic recombination: characterization of a reaction catalyzed by purified bacteriophage T4 proteins. Cell. 1986 Dec 5;47(5):793–806. doi: 10.1016/0092-8674(86)90522-2. [DOI] [PubMed] [Google Scholar]
  9. Formosa T., Burke R. L., Alberts B. M. Affinity purification of bacteriophage T4 proteins essential for DNA replication and genetic recombination. Proc Natl Acad Sci U S A. 1983 May;80(9):2442–2446. doi: 10.1073/pnas.80.9.2442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gauss P., Krassa K. B., McPheeters D. S., Nelson M. A., Gold L. Zinc (II) and the single-stranded DNA binding protein of bacteriophage T4. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8515–8519. doi: 10.1073/pnas.84.23.8515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Giedroc D. P., Keating K. M., Williams K. R., Konigsberg W. H., Coleman J. E. Gene 32 protein, the single-stranded DNA binding protein from bacteriophage T4, is a zinc metalloprotein. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8452–8456. doi: 10.1073/pnas.83.22.8452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gold L. Posttranscriptional regulatory mechanisms in Escherichia coli. Annu Rev Biochem. 1988;57:199–233. doi: 10.1146/annurev.bi.57.070188.001215. [DOI] [PubMed] [Google Scholar]
  13. Hanson J. E., Kaplan A. P., Bartlett P. A. Phosphonate analogues of carboxypeptidase A substrates are potent transition-state analogue inhibitors. Biochemistry. 1989 Jul 25;28(15):6294–6305. doi: 10.1021/bi00441a022. [DOI] [PubMed] [Google Scholar]
  14. Herendeen D. R., Kassavetis G. A., Barry J., Alberts B. M., Geiduschek E. P. Enhancement of bacteriophage T4 late transcription by components of the T4 DNA replication apparatus. Science. 1989 Sep 1;245(4921):952–958. doi: 10.1126/science.2672335. [DOI] [PubMed] [Google Scholar]
  15. Herendeen D. R., Williams K. P., Kassavetis G. A., Geiduschek E. P. An RNA polymerase-binding protein that is required for communication between an enhancer and a promoter. Science. 1990 May 4;248(4955):573–578. doi: 10.1126/science.2185541. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Jansson P. E., Kenne L., Lindberg B. Structure of extracellular polysaccharide from Xanthomonas campestris. Carbohydr Res. 1975 Dec;45:275–282. doi: 10.1016/s0008-6215(00)85885-1. [DOI] [PubMed] [Google Scholar]
  18. McPheeters D. S., Stormo G. D., Gold L. Autogenous regulatory site on the bacteriophage T4 gene 32 messenger RNA. J Mol Biol. 1988 Jun 5;201(3):517–535. doi: 10.1016/0022-2836(88)90634-1. [DOI] [PubMed] [Google Scholar]
  19. Ptashne M. How eukaryotic transcriptional activators work. Nature. 1988 Oct 20;335(6192):683–689. doi: 10.1038/335683a0. [DOI] [PubMed] [Google Scholar]
  20. Richardson J. S. The anatomy and taxonomy of protein structure. Adv Protein Chem. 1981;34:167–339. doi: 10.1016/s0065-3233(08)60520-3. [DOI] [PubMed] [Google Scholar]
  21. Sinha N. K., Morris C. F., Alberts B. M. Efficient in vitro replication of double-stranded DNA templates by a purified T4 bacteriophage replication system. J Biol Chem. 1980 May 10;255(9):4290–4293. [PubMed] [Google Scholar]
  22. Struhl K. Molecular mechanisms of transcriptional regulation in yeast. Annu Rev Biochem. 1989;58:1051–1077. doi: 10.1146/annurev.bi.58.070189.005155. [DOI] [PubMed] [Google Scholar]
  23. Stubbs G., Warren S., Holmes K. Structure of RNA and RNA binding site in tobacco mosaic virus from 4-A map calculated from X-ray fibre diagrams. Nature. 1977 May 19;267(5608):216–221. doi: 10.1038/267216a0. [DOI] [PubMed] [Google Scholar]
  24. Tan E. M. Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology. Adv Immunol. 1989;44:93–151. doi: 10.1016/s0065-2776(08)60641-0. [DOI] [PubMed] [Google Scholar]

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