Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Molecular Diversity of pagA Gene from Baciilus anthracis

탄저균 pagA 유전자의 분자적 다양성

  • 김성주 (한양대학교 생화학 및 분자생물학과) ;
  • 조기승 (한양대학교 생화학 및 분자생물학과) ;
  • 최영길 (한양대학교 생명과학과) ;
  • 채영규 (한양대학교 생화학 및 분자생물학과)
  • Published : 2001.03.01
Download PDF
⟨ Previous Next ⟩

Abstract

Bacillus anthracis is a gram-positive spore-forming bacterium that causes the disease anthrax. The anthrax toxin contains three components, including the protective antigen (PA), which binds to eucaryotic cell surface receptors and mediates the transport of toxins into the cell. In this study, the entire 2,294-nucleotide protective antigen gene (pag) was sequenced from 4 of B. anthracis strains to identify potential variation in the toxin and to further our understanding of B. anthracis evolution in Korea. Sequence alignment of the entire PA gene from 30 strains representative of the four B. anthracis diversity groups revealed mutations. The mutation of B. anthracis BAK are located adjacent to a highly antigenic region crossing the junction between PA domains 3 and 4 shown to be critical to LF binding. The different mutational combinations observed in this study give rise to 11 PA genotypes and 4PA phenotypes. Three-dimensional analysis of all the amino acid changes (Ala to Val) observed in BAK indicated that these changes are not only close sequentially but also very close in three-dimensional space to the antigenic region importan tfor LF binding. Phylogenetic (cladistic) analysis of the pag corresponded with previous strain grouping based on chromosomal variation, suggesting that plasmid evolution in B. anthracis has occurred with little or no horizontal transfer between the different strains.

탄저(anthrax)는 그람양성이고 포자형성 세균인 탄저균(Bacillus anthracis)으로부터 발명되어진다. 탄저독소는 세가지 요소로 구성되어 있으며, 방어항원(PA)은 숙주세포 표면에서 탄저 독소단백질 및 이종단백질을 세포질 내로 이동시키는 역할을 한다. 본 연구에서는 PA의 분자적 다양성과 국내에서 탄저균의 진화를 이해하고 확인하기 위해 국내외에서 발견된 탄저균 4 균주와 기존에 보고된 탄저균 26 균주로부터 2,294 bp의 PA유전자(pagA)의 DNA 염기서열을 분식하였다. 탄저균 30 균주으로부터 PA유전자의 염기서열을 비교 분식한 결과, 8개 부위에서 돌연변이를 확인 하였다. 돌연변이가 일어난 부위에 따라서 탄저균을 10종류의 PA 유전자형과 4 종류의 PA 표현형으로 구분하였다. 한국 경주에서 분류된 B. anthracis BAK는 600번째 아미노산 alanine이 valine으로 바뀌어서 B. anthracis ATCC 14185 보다 LF와 PA의 결합 위치를 근접하게 하였다. 탄저균의 Pag의 염기서열을 통한 계통분석학적인 분석 결과는 염색체상에서의 분류와 일치하여 탄저균사이에서 pXO1 플라스미드의 수평적인 이동은 없는 것으로 사료된다.

Keywords

References

  1. 한양대학교 박사학위논문 한국에서 분리한 탄저균의 분자적 다양성 연구 박경현
  2. 대한미생물학회지 v.31 탄저균 galactose-N-acetylglucosamine 다당류에 대한 단일클론 항체의 생성 및 특성 조성근;박정문;권준헌;최영길;채영규
  3. J. Bacteriol. v.178 Identification of a region of genetic variability among Bacillus anthracis strains and related species Anderson, G.L.;J.M. Simchock;K.H. Wilson
  4. Gene v.81 Nucleotide sequence and analysis of the lethal factor gene (lef) from Bacillus anthracis Bragg, T.S.;D.L. Wilson
  5. J. Clin. Microbiol. v.33 Genetic variability of Bacillus anthracis and related species Harrell, L.J.;G.L. Andersen;K.H. Wilson
  6. FEMS Microbiol. Lett. v.128 Differentiation of Bacillus anthracis and other ‘Bacillus cereus group’ bacteria using IS231-derived sequences Henderson, I.;D. Yu;P.C.B. Turnbull
  7. DNA Cloning v.Ⅱ Gene cloning in Streptomyces Hunter, I.A.;Glover D.M.(ed.)
  8. J. Bacteriol. v.179 Molecular evolution and diversity in Bacillus anthracis as detected by amplified fragment length polymorphism markers Keim, P.;A. Kalif;J. Schupp;K. Hill;S.E. Travis;K. Richmond;D.M.M. Adair;D.M.C.R. Hugh-Jones;Kuske;P. Jackson
  9. J. Appl. Microbiol. v.87 Molecular diversity in Bacillus anthracis Keim,P.;A.M. Klevytska;L.B. Price;J.M. Schupp;Zinser,G.;Smith, K.L.;M.E. Hugh-Jones;R. Okinaka;K.K. Hill;P.J. Jackson
  10. Methods Enzymol. v.165 Production and purification of anthrax toxin Leppla, S.H.
  11. J. Appl. Microbiol v.87 Identification of Bacillus anthracis strains in China Liang, X.;D.Yu
  12. Microbiology v.142 Characterization of lethal factor binding and cell receptor binding domains of protective antigen of Bacillus anthracis using monoclonal antibodies Little, S.F.;J.M. Novak;J.R. Lowe;S.H. Leppla;Y. Singh;K.R. Klimpel;B.C. Lidgerding;A.M. Friedlander
  13. Nature v.385 Crystal structure of the anthrax toxin protective antigen Petosa, C.;R.J. Collier;K.R. Klimpel;S.H. Leppla;R.C. Liddington
  14. J. Bacteriol. v.181 Genetic diversity in the protective antigen gene of Bacillus anthracis Price, L.B.;M. Hugh-Jones;P.J. Jackson;P. Protein
  15. J. Biol. Chem. v.269 The chymotrypsin-sensitive site, FFD315, in anthrax toxin protective antigen is required for translocation of lethal factor Singh, Y.;K.R. Klimpel;N. Arora;M. Sharma;S.H. Leppla
  16. Trends Microbiol. v.8 Discovery of the anthrax toxin: the beginning of in vivo studies on pathogenic bacteria Smith, H.
  17. J. Appl. Microbiol. v.87 Meso-scale ecology of anthrax in southern Africa: a pilot study of diversity and clustering Smith, K.L.;V. De;H.B. Bryden;M.E. Hugh-Jones;A. Klevytska;L.B. Price;P. Keim;D.T. Scholl
  18. Biochemistry(4th. Ed.) Stryer, L.
  19. Guidelies for the surveillance and control of anthrax in Human and Animals(3rd Ed.) Turnbull, P.C.B.;R. Bohm;O. Cosivi;M. Doganay;M.E. Hugh-Jones;D.D. Joshi;M.K. Lalitha;V. de Vos
  20. J. Bacteriol. v.175 Cloning and characterization of a gene whose product is a trans-activator of anthrax toxin synthesis Uchida, I.;J.M. Hornung;C.B. Thorne;K.R. Klimpel;S.H. Leppla
  21. Gene v.69 Sequence and analysis of the DNA encoding protective antigen of Bacillus anthracis Welkos, S.L.;J.R. Lowe;F. Eden-McCutchan;M. Vodkin;S.H. Leppla;J.J. Schmidt