Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Aug;169(8):3564–3572. doi: 10.1128/jb.169.8.3564-3572.1987

Nucleotide sequence of the Rickettsia prowazekii citrate synthase gene.

D O Wood, L R Williamson, H H Winkler, D C Krause
PMCID: PMC212433  PMID: 3112124

Abstract

The Rickettsia prowazekii citrate synthase (gltA) gene, previously cloned in Escherichia coli, was localized to a 2.0-kilobase chromosomal fragment. DNA sequence analysis of a portion of this fragment revealed an open reading frame of 1,308 base pairs that encodes a protein of 435 amino acids with a molecular weight of 49,171. This translation product is comparable in size to both the E. coli and pig heart citrate synthase monomers and to the protein synthesized in E. coli minicells containing the rickettsial gene. Comparisons between the deduced amino acid sequence of R. prowazekii citrate synthase and those of the E. coli and pig heart enzymes revealed extensive homology (59%) between the two bacterial proteins. In contrast, only 20% of the rickettsial enzyme residues were shared with the functionally similar pig heart enzyme residues. Upstream from the open reading frame and in close proximity to one another, sequences with homology to E. coli consensus sequences for RNA polymerase and ribosome binding were identified. S1 nuclease mapping experiments demonstrated that the start of transcription for this gene in E. coli was located in the upstream region. Codon usage in the rickettsial gltA gene was found to be very biased and differed from the pattern observed in E. coli. Adenine and uracil were used preferentially in the third base position of rickettsial codons.

Full text

PDF

Images in this article

3566Image
on p.3566
3569Image
on p.3569

Selected References

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

  1. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beeckmans S., Kanarek L. Purification and physicochemical characterization of chicken heart citrate synthase. Int J Biochem. 1983;15(4):469–478. doi: 10.1016/0020-711x(83)90119-2. [DOI] [PubMed] [Google Scholar]
  3. Bhayana V., Duckworth H. W. Amino acid sequence of Escherichia coli citrate synthase. Biochemistry. 1984 Jun 19;23(13):2900–2905. doi: 10.1021/bi00308a008. [DOI] [PubMed] [Google Scholar]
  4. Bloxham D. P., Parmelee D. C., Kumar S., Walsh K. A., Titani K. Complete amino acid sequence of porcine heart citrate synthase. Biochemistry. 1982 Apr 27;21(9):2028–2036. doi: 10.1021/bi00538a009. [DOI] [PubMed] [Google Scholar]
  5. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  6. Dale R. M., McClure B. A., Houchins J. P. A rapid single-stranded cloning strategy for producing a sequential series of overlapping clones for use in DNA sequencing: application to sequencing the corn mitochondrial 18 S rDNA. Plasmid. 1985 Jan;13(1):31–40. doi: 10.1016/0147-619x(85)90053-8. [DOI] [PubMed] [Google Scholar]
  7. Danson M. J., Harford S., Weitzman P. D. Studies on a mutant form of Escherichia coli citrate synthase desensitised to allosteric effectors. Eur J Biochem. 1979 Nov;101(2):515–521. doi: 10.1111/j.1432-1033.1979.tb19746.x. [DOI] [PubMed] [Google Scholar]
  8. Grantham R., Gautier C., Gouy M., Jacobzone M., Mercier R. Codon catalog usage is a genome strategy modulated for gene expressivity. Nucleic Acids Res. 1981 Jan 10;9(1):r43–r74. doi: 10.1093/nar/9.1.213-b. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hansen J. B., Abiko Y., Curtiss R., 3rd Characterization of the Streptococcus mutans plasmid pva318 cloned into Escherichia coli. Infect Immun. 1981 Mar;31(3):1034–1043. doi: 10.1128/iai.31.3.1034-1043.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Hull E. P., Spencer M. E., Wood D., Guest J. R. Nucleotide sequence of the promoter region of the citrate synthase gene (gltA) of Escherichia coli. FEBS Lett. 1983 Jun 13;156(2):366–370. doi: 10.1016/0014-5793(83)80530-4. [DOI] [PubMed] [Google Scholar]
  12. Ikemura T. Correlation between the abundance of Escherichia coli transfer RNAs and the occurrence of the respective codons in its protein genes: a proposal for a synonymous codon choice that is optimal for the E. coli translational system. J Mol Biol. 1981 Sep 25;151(3):389–409. doi: 10.1016/0022-2836(81)90003-6. [DOI] [PubMed] [Google Scholar]
  13. Krause D. C., Winkler H. H., Wood D. O. Cloning and expression of the Rickettsia prowazekii ADP/ATP translocator in Escherichia coli. Proc Natl Acad Sci U S A. 1985 May;82(9):3015–3019. doi: 10.1073/pnas.82.9.3015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Krause D. C., Winkler H. H., Wood D. O. Cosmid cloning of Rickettsia prowazekii antigens in Escherichia coli K-12. Infect Immun. 1985 Jan;47(1):157–165. doi: 10.1128/iai.47.1.157-165.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. McClure W. R. Mechanism and control of transcription initiation in prokaryotes. Annu Rev Biochem. 1985;54:171–204. doi: 10.1146/annurev.bi.54.070185.001131. [DOI] [PubMed] [Google Scholar]
  17. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  18. Mitchell C. G., Weitzman P. D. Molecular size diversity of citrate synthases from Pseudomonas species. J Gen Microbiol. 1986 Mar;132(3):737–742. doi: 10.1099/00221287-132-3-737. [DOI] [PubMed] [Google Scholar]
  19. Phibbs P. V., Jr, Winkler H. H. Regulatory properties of citrate synthase from Rickettsia prowazekii. J Bacteriol. 1982 Feb;149(2):718–725. doi: 10.1128/jb.149.2.718-725.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pribnow D. Bacteriophage T7 early promoters: nucleotide sequences of two RNA polymerase binding sites. J Mol Biol. 1975 Dec 15;99(3):419–443. doi: 10.1016/s0022-2836(75)80136-7. [DOI] [PubMed] [Google Scholar]
  21. Pribnow D. Nucleotide sequence of an RNA polymerase binding site at an early T7 promoter. Proc Natl Acad Sci U S A. 1975 Mar;72(3):784–788. doi: 10.1073/pnas.72.3.784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Remington S., Wiegand G., Huber R. Crystallographic refinement and atomic models of two different forms of citrate synthase at 2.7 and 1.7 A resolution. J Mol Biol. 1982 Jun 15;158(1):111–152. doi: 10.1016/0022-2836(82)90452-1. [DOI] [PubMed] [Google Scholar]
  23. Rhoads D. B., Laimins L., Epstein W. Functional organization of the kdp genes of Escherichia coli K-12. J Bacteriol. 1978 Aug;135(2):445–452. doi: 10.1128/jb.135.2.445-452.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Robeson J. P., Barletta R. G., Curtiss R., 3rd Expression of a Streptococcus mutans glucosyltransferase gene in Escherichia coli. J Bacteriol. 1983 Jan;153(1):211–221. doi: 10.1128/jb.153.1.211-221.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Robinson M. S., Danson M. J., Weitzman P. D. Citrate synthase from a Gram-positive bacterium. Purification and characterization of the Bacillus megaterium enzyme. Biochem J. 1983 Jul 1;213(1):53–59. doi: 10.1042/bj2130053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Seeburg P. H., Nüsslein C., Schaller H. Interaction of RNA polymerase with promoters from bacteriophage fd. Eur J Biochem. 1977 Mar 15;74(1):107–113. doi: 10.1111/j.1432-1033.1977.tb11372.x. [DOI] [PubMed] [Google Scholar]
  29. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Suissa M., Suda K., Schatz G. Isolation of the nuclear yeast genes for citrate synthase and fifteen other mitochondrial proteins by a new screening method. EMBO J. 1984 Aug;3(8):1773–1781. doi: 10.1002/j.1460-2075.1984.tb02045.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  33. Weiss E. Growth and physiology of rickettsiae. Bacteriol Rev. 1973 Sep;37(3):259–283. doi: 10.1128/br.37.3.259-283.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weiss E. The biology of rickettsiae. Annu Rev Microbiol. 1982;36:345–370. doi: 10.1146/annurev.mi.36.100182.002021. [DOI] [PubMed] [Google Scholar]
  35. Weitzman P. D., Danson M. J. Citrate synthase. Curr Top Cell Regul. 1976;10:161–204. doi: 10.1016/b978-0-12-152810-2.50011-5. [DOI] [PubMed] [Google Scholar]
  36. West R. W., Jr, Rodriguez R. L. Construction and characterization of E. coli promoter-probe plasmid vectors. III. pBR322 derivatives with deletions in the tetracycline resistance promoter region. Gene. 1982 Dec;20(2):291–304. doi: 10.1016/0378-1119(82)90047-6. [DOI] [PubMed] [Google Scholar]
  37. Wiegand G., Remington S. J. Citrate synthase: structure, control, and mechanism. Annu Rev Biophys Biophys Chem. 1986;15:97–117. doi: 10.1146/annurev.bb.15.060186.000525. [DOI] [PubMed] [Google Scholar]
  38. Winkler H. H. Rickettsial permeability. An ADP-ATP transport system. J Biol Chem. 1976 Jan 25;251(2):389–396. [PubMed] [Google Scholar]
  39. Wood D. O., Atkinson W. H., Sikorski R. S., Winkler H. H. Expression of the Rickettsia prowazekii citrate synthase gene in Escherichia coli. J Bacteriol. 1983 Jul;155(1):412–416. doi: 10.1128/jb.155.1.412-416.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES