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. 1988 Jul;56(7):1678–1684. doi: 10.1128/iai.56.7.1678-1684.1988

Developmental-form-specific DNA-binding proteins in Chlamydia spp.

E A Wagar 1, R S Stephens 1
PMCID: PMC259462  PMID: 3384472

Abstract

We identified DNA-binding proteins specific to the elementary body (EB) developmental form of Chlamydia spp. Chlamydial proteins from whole lysates were separated by polyacrylamide gel electrophoresis, transferred to nitrocellulose, and probed with nick-translated chlamydial DNA. By this method, C. trachomatis serovar L2 EBs had three unique protein bands of 58,000, 25,700, and 17,000 molecular weight not seen in the reticulate bodies. The 17,000-molecular-weight protein and the 25,700-molecular-weight protein were identified in an acid-soluble protein fraction and were resistant to high-salt elution, similar to other procaryotic nucleoproteins. The 17,000-molecular-weight protein was also detected in preparations with isolated chromosomes from EBs. Preliminary characterization indicated that the protein-DNA interaction was principally charge related.

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Selected References

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

  1. Batteiger B. E., Newhall W. J., 5th, Jones R. B. Differences in outer membrane proteins of the lymphogranuloma venereum and trachoma biovars of Chlamydia trachomatis. Infect Immun. 1985 Nov;50(2):488–494. doi: 10.1128/iai.50.2.488-494.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bowen B., Steinberg J., Laemmli U. K., Weintraub H. The detection of DNA-binding proteins by protein blotting. Nucleic Acids Res. 1980 Jan 11;8(1):1–20. doi: 10.1093/nar/8.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Broyles S. S., Pettijohn D. E. Interaction of the Escherichia coli HU protein with DNA. Evidence for formation of nucleosome-like structures with altered DNA helical pitch. J Mol Biol. 1986 Jan 5;187(1):47–60. doi: 10.1016/0022-2836(86)90405-5. [DOI] [PubMed] [Google Scholar]
  4. Bujalowski W., Lohman T. M. Escherichia coli single-strand binding protein forms multiple, distinct complexes with single-stranded DNA. Biochemistry. 1986 Dec 2;25(24):7799–7802. doi: 10.1021/bi00372a003. [DOI] [PubMed] [Google Scholar]
  5. Caldwell H. D., Kromhout J., Schachter J. Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis. Infect Immun. 1981 Mar;31(3):1161–1176. doi: 10.1128/iai.31.3.1161-1176.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Costerton J. W., Poffenroth L., Wilt J. C., Kordová N. Ultrastructural studies of the nucleoids of the pleomorphic forms of Chlamydia psittaci 6BC: a comparison with bacteria. Can J Microbiol. 1976 Jan;22(1):16–28. doi: 10.1139/m76-003. [DOI] [PubMed] [Google Scholar]
  7. Drlica K., Rouviere-Yaniv J. Histonelike proteins of bacteria. Microbiol Rev. 1987 Sep;51(3):301–319. doi: 10.1128/mr.51.3.301-319.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gicquel-Sanzey B., Cossart P. Homologies between different procaryotic DNA-binding regulatory proteins and between their sites of action. EMBO J. 1982;1(5):591–595. doi: 10.1002/j.1460-2075.1982.tb01213.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grayston J. T., Kuo C. C., Wang S. P., Altman J. A new Chlamydia psittaci strain, TWAR, isolated in acute respiratory tract infections. N Engl J Med. 1986 Jul 17;315(3):161–168. doi: 10.1056/NEJM198607173150305. [DOI] [PubMed] [Google Scholar]
  10. Hackstadt T. Identification and properties of chlamydial polypeptides that bind eucaryotic cell surface components. J Bacteriol. 1986 Jan;165(1):13–20. doi: 10.1128/jb.165.1.13-20.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hackstadt T., Todd W. J., Caldwell H. D. Disulfide-mediated interactions of the chlamydial major outer membrane protein: role in the differentiation of chlamydiae? J Bacteriol. 1985 Jan;161(1):25–31. doi: 10.1128/jb.161.1.25-31.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hatch T. P., Allan I., Pearce J. H. Structural and polypeptide differences between envelopes of infective and reproductive life cycle forms of Chlamydia spp. J Bacteriol. 1984 Jan;157(1):13–20. doi: 10.1128/jb.157.1.13-20.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hübscher U., Lutz H., Kornberg A. Novel histone H2A-like protein of escherichia coli. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5097–5101. doi: 10.1073/pnas.77.9.5097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kano Y., Wada M., Nagase T., Imamoto F. Genetic characterization of the gene hupB encoding the HU-1 protein of Escherichia coli. Gene. 1986;45(1):37–44. doi: 10.1016/0378-1119(86)90129-0. [DOI] [PubMed] [Google Scholar]
  15. Kimura M., Wilson K. S. On the DNA binding protein II from Bacillus stearothermophilus. II. The amino acid sequence and its relation to those of homologous proteins from other prokaryotes. J Biol Chem. 1983 Mar 25;258(6):4007–4011. [PubMed] [Google Scholar]
  16. 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]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Laine B., Bélaïche D., Khanaka H., Sautière P. Primary structure of the DNA-binding protein HRm from Rhizobium meliloti. Eur J Biochem. 1983 Mar 15;131(2):325–331. doi: 10.1111/j.1432-1033.1983.tb07265.x. [DOI] [PubMed] [Google Scholar]
  19. Lathe R., Buc H., Lecocq J. P., Bautz E. K. Prokaryotic histone-like protein interacting with RNA polymerase. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3548–3552. doi: 10.1073/pnas.77.6.3548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Newhall W. J., 5th Biosynthesis and disulfide cross-linking of outer membrane components during the growth cycle of Chlamydia trachomatis. Infect Immun. 1987 Jan;55(1):162–168. doi: 10.1128/iai.55.1.162-168.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  22. Pettijohn D. E. Structure and properties of the bacterial nucleoid. Cell. 1982 Oct;30(3):667–669. doi: 10.1016/0092-8674(82)90269-0. [DOI] [PubMed] [Google Scholar]
  23. Popov V., Eb F., Lefebvre J. F., Orfila J., Viron A. Morphological and cytochemical study of Chlamydia with EDTA regressive technique and Gautier staining in ultrathin frozen sections of infected cell cultures: a comparison with embedded material. Ann Microbiol (Paris) 1978 Oct;129 B(3):313–337. [PubMed] [Google Scholar]
  24. Rouvière-Yaniv J., Gros F. Characterization of a novel, low-molecular-weight DNA-binding protein from Escherichia coli. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3428–3432. doi: 10.1073/pnas.72.9.3428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schachter J., Caldwell H. D. Chlamydiae. Annu Rev Microbiol. 1980;34:285–309. doi: 10.1146/annurev.mi.34.100180.001441. [DOI] [PubMed] [Google Scholar]
  26. Schachter J. Chlamydial infections (third of three parts). N Engl J Med. 1978 Mar 9;298(10):540–549. doi: 10.1056/NEJM197803092981005. [DOI] [PubMed] [Google Scholar]
  27. Searcy D. G. Histone-like protein in the prokaryote Thermoplasma acidophilum. Biochim Biophys Acta. 1975 Jul 23;395(4):535–547. doi: 10.1016/0005-2787(75)90076-3. [DOI] [PubMed] [Google Scholar]
  28. Silva C. M., Tully D. B., Petch L. A., Jewell C. M., Cidlowski J. A. Application of a protein-blotting procedure to the study of human glucocorticoid receptor interactions with DNA. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1744–1748. doi: 10.1073/pnas.84.7.1744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sjåstad K., Fadnes P., Krüger P. G., Lossius I., Kleppe K. Isolation, properties and nucleolytic degradation of chromatin from Escherichia coli. J Gen Microbiol. 1982 Dec;128(12):3037–3050. doi: 10.1099/00221287-128-12-3037. [DOI] [PubMed] [Google Scholar]
  30. Spassky A., Rimsky S., Garreau H., Buc H. H1a, an E. coli DNA-binding protein which accumulates in stationary phase, strongly compacts DNA in vitro. Nucleic Acids Res. 1984 Jul 11;12(13):5321–5340. doi: 10.1093/nar/12.13.5321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Stephens R. S., Kuo C. C., Newport G., Agabian N. Molecular cloning and expression of Chlamydia trachomatis major outer membrane protein antigens in Escherichia coli. Infect Immun. 1985 Mar;47(3):713–718. doi: 10.1128/iai.47.3.713-718.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sutrina S. L., Scocca J. J. DNA-binding proteins of Haemophilus influenzae: purification and characterization of a major intracellular binding protein. J Bacteriol. 1983 Jul;155(1):246–253. doi: 10.1128/jb.155.1.246-253.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weisburg W. G., Hatch T. P., Woese C. R. Eubacterial origin of chlamydiae. J Bacteriol. 1986 Aug;167(2):570–574. doi: 10.1128/jb.167.2.570-574.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wenman W. M., Meuser R. U. Chlamydia trachomatis elementary bodies possess proteins which bind to eucaryotic cell membranes. J Bacteriol. 1986 Feb;165(2):602–607. doi: 10.1128/jb.165.2.602-607.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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