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. 1990 Sep;58(9):2760–2769. doi: 10.1128/iai.58.9.2760-2769.1990

A protective protein antigen of Rickettsia rickettsii has tandemly repeated, near-identical sequences.

B E Anderson 1, G A McDonald 1, D C Jones 1, R L Regnery 1
PMCID: PMC313565  PMID: 2117568

Abstract

The nucleotide sequence of a Rickettsia rickettsii gene that encodes a high-molecular-mass surface antigen (190 kilodaltons), which elicits protective immunity, was determined. The 6,747-nucleotide gene coded for a 2,249-amino-acid protein with a calculated molecular weight of 224,321. A 3.8-kilobase PstI fragment proximal to the 5' end of the gene was found to consist of 13 highly related tandem repeats which constituted over 40% of the coding region. The repeated sequences could be divided into either a 225-nucleotide, 75-amino-acid unit (type I) or a 216-nucleotide, 72-amino-acid unit (type II), with extensive homology between the two types of repeating units. The deduced amino acid sequence for these repeat units, overall, was slightly hydrophobic with short hydrophilic domains. The carboxy-terminal (nonrepetitive) portion of the deduced protein sequence was hydrophilic, with potential surface-exposed epitopes. The full-length reading frame was reconstructed in Escherichia coli, and transient expression of the 190-kilodalton antigen was demonstrated; however, the protein appeared to be severely degraded by proteases and was apparently toxic to E. coli. The conservation of this unique repetitive gene structure, coupled with results from previous reports showing the protective properties of the 190-kilodalton antigen, suggests that this protein plays an important role in the pathogenesis of and immunity to Rocky Mountain spotted fever.

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Selected References

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  1. Anacker R. L., List R. H., Mann R. E., Hayes S. F., Thomas L. A. Characterization of monoclonal antibodies protecting mice against Rickettsia rickettsii. J Infect Dis. 1985 Jun;151(6):1052–1060. doi: 10.1093/infdis/151.6.1052. [DOI] [PubMed] [Google Scholar]
  2. Anacker R. L., List R. H., Mann R. E., Wiedbrauk D. L. Antigenic heterogeneity in high- and low-virulence strains of Rickettsia rickettsii revealed by monoclonal antibodies. Infect Immun. 1986 Feb;51(2):653–660. doi: 10.1128/iai.51.2.653-660.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anacker R. L., Mann R. E., Gonzales C. Reactivity of monoclonal antibodies to Rickettsia rickettsii with spotted fever and typhus group rickettsiae. J Clin Microbiol. 1987 Jan;25(1):167–171. doi: 10.1128/jcm.25.1.167-171.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anacker R. L., McDonald G. A., List R. H., Mann R. E. Neutralizing activity of monoclonal antibodies to heat-sensitive and heat-resistant epitopes of Rickettsia rickettsii surface proteins. Infect Immun. 1987 Mar;55(3):825–827. doi: 10.1128/iai.55.3.825-827.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Anders R. F. Multiple cross-reactivities amongst antigens of Plasmodium falciparum impair the development of protective immunity against malaria. Parasite Immunol. 1986 Nov;8(6):529–539. doi: 10.1111/j.1365-3024.1986.tb00867.x. [DOI] [PubMed] [Google Scholar]
  6. Anderson B. E., Baumstark B. R., Bellini W. J. Expression of the gene encoding the 17-kilodalton antigen from Rickettsia rickettsii: transcription and posttranslational modification. J Bacteriol. 1988 Oct;170(10):4493–4500. doi: 10.1128/jb.170.10.4493-4500.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Anderson B. E., Regnery R. L., Carlone G. M., Tzianabos T., McDade J. E., Fu Z. Y., Bellini W. J. Sequence analysis of the 17-kilodalton-antigen gene from Rickettsia rickettsii. J Bacteriol. 1987 Jun;169(6):2385–2390. doi: 10.1128/jb.169.6.2385-2390.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Anderson B. E., Tzianabos T. Comparative sequence analysis of a genus-common rickettsial antigen gene. J Bacteriol. 1989 Sep;171(9):5199–5201. doi: 10.1128/jb.171.9.5199-5201.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. BELL E. J., PICKENS E. G. A toxic substance associated with the rickettsias of the spotted fever group. J Immunol. 1953 May;70(5):461–472. [PubMed] [Google Scholar]
  10. Büchel D. E., Gronenborn B., Müller-Hill B. Sequence of the lactose permease gene. Nature. 1980 Feb 7;283(5747):541–545. doi: 10.1038/283541a0. [DOI] [PubMed] [Google Scholar]
  11. Clements M. L., Wisseman C. L., Jr, Woodward T. E., Fiset P., Dumler J. S., McNamee W., Black R. E., Rooney J., Hughes T. P., Levine M. M. Reactogenicity, immunogenicity, and efficacy of a chick embryo cell-derived vaccine for Rocky Mountain spotted fever. J Infect Dis. 1983 Nov;148(5):922–930. doi: 10.1093/infdis/148.5.922. [DOI] [PubMed] [Google Scholar]
  12. DuPont H. L., Hornick R. B., Dawkins A. T., Heiner G. G., Fabrikant I. B., Wisseman C. L., Jr, Woodward T. E. Rocky Mountain spotted fever: a comparative study of the active immunity induced by inactivated and viable pathogenic Rickettsia rickettsii. J Infect Dis. 1973 Sep;128(3):340–344. doi: 10.1093/infdis/128.3.340. [DOI] [PubMed] [Google Scholar]
  13. Dubreuil J. D., Logan S. M., Cubbage S., Eidhin D. N., McCubbin W. D., Kay C. M., Beveridge T. J., Ferris F. G., Trust T. J. Structural and biochemical analyses of a surface array protein of Campylobacter fetus. J Bacteriol. 1988 Sep;170(9):4165–4173. doi: 10.1128/jb.170.9.4165-4173.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gilmore R. D., Jr, Joste N., McDonald G. A. Cloning, expression and sequence analysis of the gene encoding the 120 kD surface-exposed protein of Rickettsia rickettsii. Mol Microbiol. 1989 Nov;3(11):1579–1586. doi: 10.1111/j.1365-2958.1989.tb00143.x. [DOI] [PubMed] [Google Scholar]
  15. Hasan N., Szybalski W. Control of cloned gene expression by promoter inversion in vivo: construction of improved vectors with a multiple cloning site and the Ptac promoter. Gene. 1987;56(1):145–151. doi: 10.1016/0378-1119(87)90167-3. [DOI] [PubMed] [Google Scholar]
  16. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hoft D. F., Kim K. S., Otsu K., Moser D. R., Yost W. J., Blumin J. H., Donelson J. E., Kirchhoff L. V. Trypanosoma cruzi expresses diverse repetitive protein antigens. Infect Immun. 1989 Jul;57(7):1959–1967. doi: 10.1128/iai.57.7.1959-1967.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ibañez C. F., Affranchino J. L., Macina R. A., Reyes M. B., Leguizamon S., Camargo M. E., Aslund L., Pettersson U., Frasch A. C. Multiple Trypanosoma cruzi antigens containing tandemly repeated amino acid sequence motifs. Mol Biochem Parasitol. 1988 Jul;30(1):27–33. doi: 10.1016/0166-6851(88)90129-6. [DOI] [PubMed] [Google Scholar]
  19. Ito K. Identification of the secY (prlA) gene product involved in protein export in Escherichia coli. Mol Gen Genet. 1984;197(2):204–208. doi: 10.1007/BF00330964. [DOI] [PubMed] [Google Scholar]
  20. Jerrells T. R., Jarboe D. L., Eisemann C. S. Cross-reactive lymphocyte responses and protective immunity against other spotted fever group rickettsiae in mice immunized with Rickettsia conorii. Infect Immun. 1986 Mar;51(3):832–837. doi: 10.1128/iai.51.3.832-837.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kemp D. J., Coppel R. L., Stahl H. D., Bianco A. E., Corcoran L. M., McIntyre P., Langford C. J., Favaloro J. M., Crewther P. E., Brown G. V. The Wellcome Trust lecture. Genes for antigens of Plasmodium falciparum. Parasitology. 1986;92 (Suppl):S83–108. doi: 10.1017/s0031182000085711. [DOI] [PubMed] [Google Scholar]
  22. Kenyon R. H., Ascher M. S., Kishimoto R. A., Pedersen C. E., Jr In vitro guinea pig leukocyte reactions to Rickettsia rickettsii. Infect Immun. 1977 Dec;18(3):840–846. doi: 10.1128/iai.18.3.840-846.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kokorin I. N., Kabanova E. A., Shirokova E. M., Abrosimova G. E., Rybkina N. N., Pushkareva V. i. Role of T lymphocytes in Rickettsia conorii infection. Acta Virol. 1982 Jan;26(1-2):91–97. [PubMed] [Google Scholar]
  24. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Lechner J., Sumper M. The primary structure of a procaryotic glycoprotein. Cloning and sequencing of the cell surface glycoprotein gene of halobacteria. J Biol Chem. 1987 Jul 15;262(20):9724–9729. [PubMed] [Google Scholar]
  27. Marchalonis J. J. An enzymic method for the trace iodination of immunoglobulins and other proteins. Biochem J. 1969 Jun;113(2):299–305. doi: 10.1042/bj1130299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mazier D., Mellouk S., Beaudoin R. L., Texier B., Druilhe P., Hockmeyer W., Trosper J., Paul C., Charoenvit Y., Young J. Effect of antibodies to recombinant and synthetic peptides on P. falciparum sporozoites in vitro. Science. 1986 Jan 10;231(4734):156–159. doi: 10.1126/science.3510455. [DOI] [PubMed] [Google Scholar]
  29. McDonald G. A., Anacker R. L., Garjian K. Cloned gene of Rickettsia rickettsii surface antigen: candidate vaccine for Rocky Mountain spotted fever. Science. 1987 Jan 2;235(4784):83–85. doi: 10.1126/science.3099387. [DOI] [PubMed] [Google Scholar]
  30. McDonald G. A., Anacker R. L., Mann R. E., Milch L. J. Protection of guinea pigs from experimental Rocky Mountain spotted fever with a cloned antigen of Rickettsia rickettsii. J Infect Dis. 1988 Jul;158(1):228–231. doi: 10.1093/infdis/158.1.228. [DOI] [PubMed] [Google Scholar]
  31. Nicaud J. M., Mackman N., Gray L., Holland I. B. The C-terminal, 23 kDa peptide of E. coli haemolysin 2001 contains all the information necessary for its secretion by the haemolysin (Hly) export machinery. FEBS Lett. 1986 Aug 18;204(2):331–335. doi: 10.1016/0014-5793(86)80838-9. [DOI] [PubMed] [Google Scholar]
  32. Palmer E. L., Mallavia L. P., Tzianabos T., Obijeski J. F. Electron microscopy of the cell wall of Rickettsia prowazeki. J Bacteriol. 1974 Jun;118(3):1158–1166. doi: 10.1128/jb.118.3.1158-1166.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Palmer E. L., Martin M. L., Mallavia L. Ultrastucture of the surface of Rickettsia prowazeki and Rickettsia akari. Appl Microbiol. 1974 Oct;28(4):713–716. doi: 10.1128/am.28.4.713-716.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Roditi I., Carrington M., Turner M. Expression of a polypeptide containing a dipeptide repeat is confined to the insect stage of Trypanosoma brucei. Nature. 1987 Jan 15;325(6101):272–274. doi: 10.1038/325272a0. [DOI] [PubMed] [Google Scholar]
  35. Schneider A., Hemphill A., Wyler T., Seebeck T. Large microtubule-associated protein of T. brucei has tandemly repeated, near-identical sequences. Science. 1988 Jul 22;241(4864):459–462. doi: 10.1126/science.3393912. [DOI] [PubMed] [Google Scholar]
  36. Silhavy T. J., Benson S. A., Emr S. D. Mechanisms of protein localization. Microbiol Rev. 1983 Sep;47(3):313–344. doi: 10.1128/mr.47.3.313-344.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sleytr U. B., Messner P. Crystalline surface layers in procaryotes. J Bacteriol. 1988 Jul;170(7):2891–2897. doi: 10.1128/jb.170.7.2891-2897.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sleytr U. B., Messner P. Crystalline surface layers on bacteria. Annu Rev Microbiol. 1983;37:311–339. doi: 10.1146/annurev.mi.37.100183.001523. [DOI] [PubMed] [Google Scholar]
  39. Stark M. J. Multicopy expression vectors carrying the lac repressor gene for regulated high-level expression of genes in Escherichia coli. Gene. 1987;51(2-3):255–267. doi: 10.1016/0378-1119(87)90314-3. [DOI] [PubMed] [Google Scholar]
  40. Stormo G. D., Schneider T. D., Gold L. M. Characterization of translational initiation sites in E. coli. Nucleic Acids Res. 1982 May 11;10(9):2971–2996. doi: 10.1093/nar/10.9.2971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. Walker D. H., Montenegro M. R., Hegarty B. C., Tringali G. R. Rocky Mountain spotted fever vaccine: a regional need. South Med J. 1984 Apr;77(4):447–449. doi: 10.1097/00007611-198404000-00009. [DOI] [PubMed] [Google Scholar]
  43. Wallis A. E., McMaster W. R. Identification of Leishmania genes encoding proteins containing tandemly repeating peptides. J Exp Med. 1987 Dec 1;166(6):1814–1824. doi: 10.1084/jem.166.6.1814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Weiss E., Coolbaugh J. C., Williams J. C. Separation of viable Rickettsia typhi from yolk sac and L cell host components by renografin density gradient centrifugation. Appl Microbiol. 1975 Sep;30(3):456–463. doi: 10.1128/am.30.3.456-463.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Williams J. C., Walker D. H., Peacock M. G., Stewart S. T. Humoral immune response to Rocky Mountain spotted fever in experimentally infected guinea pigs: immunoprecipitation of lactoperoxidase 125I-labeled proteins and detection of soluble antigens of Rickettsia rickettsii. Infect Immun. 1986 Apr;52(1):120–127. doi: 10.1128/iai.52.1.120-127.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Williamson L. R., Plano G. V., Winkler H. H., Krause D. C., Wood D. O. Nucleotide sequence of the Rickettsia prowazekii ATP/ADP translocase-encoding gene. Gene. 1989 Aug 15;80(2):269–278. doi: 10.1016/0378-1119(89)90291-6. [DOI] [PubMed] [Google Scholar]
  47. Wood D. O., Williamson L. R., Winkler H. H., Krause D. C. Nucleotide sequence of the Rickettsia prowazekii citrate synthase gene. J Bacteriol. 1987 Aug;169(8):3564–3572. doi: 10.1128/jb.169.8.3564-3572.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  49. Young J. F., Hockmeyer W. T., Gross M., Ballou W. R., Wirtz R. A., Trosper J. H., Beaudoin R. L., Hollingdale M. R., Miller L. H., Diggs C. L. Expression of Plasmodium falciparum circumsporozoite proteins in Escherichia coli for potential use in a human malaria vaccine. Science. 1985 May 24;228(4702):958–962. doi: 10.1126/science.2988125. [DOI] [PubMed] [Google Scholar]
  50. Zhang H., Scholl R., Browse J., Somerville C. Double stranded DNA sequencing as a choice for DNA sequencing. Nucleic Acids Res. 1988 Feb 11;16(3):1220–1220. doi: 10.1093/nar/16.3.1220. [DOI] [PMC free article] [PubMed] [Google Scholar]

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