Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1991 Aug;59(8):2695–2705. doi: 10.1128/iai.59.8.2695-2705.1991

Insertion element IS987 from Mycobacterium bovis BCG is located in a hot-spot integration region for insertion elements in Mycobacterium tuberculosis complex strains.

P W Hermans 1, D van Soolingen 1, E M Bik 1, P E de Haas 1, J W Dale 1, J D van Embden 1
PMCID: PMC258075  PMID: 1649798

Abstract

Most strains of the Mycobacterium tuberculosis complex carry multiple copies of an IS3-like element, and these strains are highly polymorphic with regard to the site of integration in the chromosome. In contrast, Mycobacterium bovis BCG contains a single copy of the insertion element, and in all strains this copy is integrated at the same site in the chromosome. In this study, we determined the sequence of the single-copy insertion element from M. bovis BCG, IS987, and its flanking regions. The analysis of IS987 revealed that this element was virtually identical to the sequence of IS986 from M. tuberculosis. IS987 is located in a region containing direct repeats (DRs). The cloned flanking regions contained 20 virtually identical DRs of 36 bp, each separated by 35 to 41 bp of spacer DNA. Analysis of chromosomal DNA by the polymerase chain reaction revealed the presence of a cluster of 49 DRs, and IS987 is inserted in the 30th DR. Furthermore, the DR sequences were found to occur only in species of the M. tuberculosis complex and not in nine other mycobacterial species tested. Analysis of 14 M. tuberculosis strains revealed the presence of one insertion sequence element in the DR-containing region of eight strains, two insertion sequence elements were located in the DR region of five strains, and one strain did not contain an insertion sequence element in this region. Additionally, the DR-containing regions of these 14 M. tuberculosis strains were polymorphic in length and composition. We conclude that the DR cluster is a specific, hot-spot region for integration of insertion elements in the chromosome of M. tuberculosis complex strains.

Full text

PDF
2695

Images in this article

2701Image
on p.2701
2701Image
on p.2701
2702Image
on p.2702
2703Image
on p.2703
2703Image
on p.2703
2704Image
on p.2704

Selected References

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

  1. Barberis-Maino L., Berger-Bächi B., Weber H., Beck W. D., Kayser F. H. IS431, a staphylococcal insertion sequence-like element related to IS26 from Proteus vulgaris. Gene. 1987;59(1):107–113. doi: 10.1016/0378-1119(87)90271-x. [DOI] [PubMed] [Google Scholar]
  2. Campbell A., Starlinger P., Berg D. E., Botstein D., Lederberg E. M., Novick R. P., Szybalski W. Nomenclature of transposable elements in prokaryotes. Plasmid. 1979 Jul;2(3):466–473. doi: 10.1016/0147-619x(79)90030-1. [DOI] [PubMed] [Google Scholar]
  3. Green E. P., Tizard M. L., Moss M. T., Thompson J., Winterbourne D. J., McFadden J. J., Hermon-Taylor J. Sequence and characteristics of IS900, an insertion element identified in a human Crohn's disease isolate of Mycobacterium paratuberculosis. Nucleic Acids Res. 1989 Nov 25;17(22):9063–9073. doi: 10.1093/nar/17.22.9063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hampson S. J., Portaels F., Thompson J., Green E. P., Moss M. T., Hermon-Taylor J., McFadden J. J. DNA probes demonstrate a single highly conserved strain of Mycobacterium avium infecting AIDS patients. Lancet. 1989 Jan 14;1(8629):65–68. doi: 10.1016/s0140-6736(89)91427-x. [DOI] [PubMed] [Google Scholar]
  5. Hermans P. W., Schuitema A. R., Van Soolingen D., Verstynen C. P., Bik E. M., Thole J. E., Kolk A. H., van Embden J. D. Specific detection of Mycobacterium tuberculosis complex strains by polymerase chain reaction. J Clin Microbiol. 1990 Jun;28(6):1204–1213. doi: 10.1128/jcm.28.6.1204-1213.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hermans P. W., van Soolingen D., Dale J. W., Schuitema A. R., McAdam R. A., Catty D., van Embden J. D. Insertion element IS986 from Mycobacterium tuberculosis: a useful tool for diagnosis and epidemiology of tuberculosis. J Clin Microbiol. 1990 Sep;28(9):2051–2058. doi: 10.1128/jcm.28.9.2051-2058.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ishiguro N., Sato G. Spontaneous deletion of citrate-utilizing ability promoted by insertion sequences. J Bacteriol. 1984 Nov;160(2):642–650. doi: 10.1128/jb.160.2.642-650.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jordan E., Saedler H., Starlinger P. O0 and strong-polar mutations in the gal operon are insertions. Mol Gen Genet. 1968;102(4):353–363. doi: 10.1007/BF00433726. [DOI] [PubMed] [Google Scholar]
  9. Mahillon J., Seurinck J., van Rompuy L., Delcour J., Zabeau M. Nucleotide sequence and structural organization of an insertion sequence element (IS231) from Bacillus thuringiensis strain berliner 1715. EMBO J. 1985 Dec 30;4(13B):3895–3899. doi: 10.1002/j.1460-2075.1985.tb04163.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Martin C., Timm J., Rauzier J., Gomez-Lus R., Davies J., Gicquel B. Transposition of an antibiotic resistance element in mycobacteria. Nature. 1990 Jun 21;345(6277):739–743. doi: 10.1038/345739a0. [DOI] [PubMed] [Google Scholar]
  11. Matsutani S., Ohtsubo H., Maeda Y., Ohtsubo E. Isolation and characterization of IS elements repeated in the bacterial chromosome. J Mol Biol. 1987 Aug 5;196(3):445–455. doi: 10.1016/0022-2836(87)90023-4. [DOI] [PubMed] [Google Scholar]
  12. McAdam R. A., Hermans P. W., van Soolingen D., Zainuddin Z. F., Catty D., van Embden J. D., Dale J. W. Characterization of a Mycobacterium tuberculosis insertion sequence belonging to the IS3 family. Mol Microbiol. 1990 Sep;4(9):1607–1613. doi: 10.1111/j.1365-2958.1990.tb02073.x. [DOI] [PubMed] [Google Scholar]
  13. McFadden J. J., Butcher P. D., Thompson J., Chiodini R., Hermon-Taylor J. The use of DNA probes identifying restriction-fragment-length polymorphisms to examine the Mycobacterium avium complex. Mol Microbiol. 1987 Nov;1(3):283–291. doi: 10.1111/j.1365-2958.1987.tb01934.x. [DOI] [PubMed] [Google Scholar]
  14. Nash J. H., Krishnapillai V. Identification of an insertion sequence in the chromosome of Pseudomonas aeruginosa PAO. J Bacteriol. 1982 Oct;152(1):514–516. doi: 10.1128/jb.152.1.514-516.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ooms G., Hooykaas P. J., Moolenaar G., Schilperoort R. A. Grown gall plant tumors of abnormal morphology, induced by Agrobacterium tumefaciens carrying mutated octopine Ti plasmids; analysis of T-DNA functions. Gene. 1981 Jun-Jul;14(1-2):33–50. doi: 10.1016/0378-1119(81)90146-3. [DOI] [PubMed] [Google Scholar]
  16. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Priefer U. B., Burkardt H. J., Klipp W., Pühler A. ISR1: an insertion element isolated from the soil bacterium Rhizobium lupini. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):87–91. doi: 10.1101/sqb.1981.045.01.016. [DOI] [PubMed] [Google Scholar]
  18. Robinson A., Ashworth L. A., Baskerville A., Irons L. I. Protection against intranasal infection of mice with Bordetella pertussis. Dev Biol Stand. 1985;61:165–172. [PubMed] [Google Scholar]
  19. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Thierry D., Brisson-Noël A., Vincent-Lévy-Frébault V., Nguyen S., Guesdon J. L., Gicquel B. Characterization of a Mycobacterium tuberculosis insertion sequence, IS6110, and its application in diagnosis. J Clin Microbiol. 1990 Dec;28(12):2668–2673. doi: 10.1128/jcm.28.12.2668-2673.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Thole J. E., Keulen W. J., De Bruyn J., Kolk A. H., Groothuis D. G., Berwald L. G., Tiesjema R. H., van Embden J. D. Characterization, sequence determination, and immunogenicity of a 64-kilodalton protein of Mycobacterium bovis BCG expressed in escherichia coli K-12. Infect Immun. 1987 Jun;55(6):1466–1475. doi: 10.1128/iai.55.6.1466-1475.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Zainuddin Z. F., Dale J. W. Polymorphic repetitive DNA sequences in Mycobacterium tuberculosis detected with a gene probe from a Mycobacterium fortuitum plasmid. J Gen Microbiol. 1989 Sep;135(9):2347–2355. doi: 10.1099/00221287-135-9-2347. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES