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. 1991 Dec;173(24):7802–7809. doi: 10.1128/jb.173.24.7802-7809.1991

Multiple copies of IS10 in the Enterobacter cloacae MD36 chromosome.

S Matsutani 1
PMCID: PMC212570  PMID: 1660455

Abstract

Repetitive sequences were isolated and characterized as double-stranded DNA fragments by treatment with S1 nuclease after denaturation and renaturation of the total DNA of Enterobacter cloacae MD36. One repetitive sequence was identical to the nucleotide sequence of IS10-right (IS10R), which is the active element in the plasmid-associated transposon Tn10. Unexpectedly, 15 copies of IS10R were found in the chromosomal DNA of E. cloacae MD36. One copy of the central region of Tn10 was found in the total DNA of E. cloacae MD36. IS10Rs in restriction fragments isolated from the E. cloacae MD36 total DNA showed 9-bp duplications adjacent to the terminal sequences that are characteristic of Tn10 transposition. This result suggests that many copies of IS10R in E. cloacae MD36 are due to transposition of IS10R alone, not due to transposition of Tn10 or to DNA rearrangement. I also found nine copies of IS10 in Shigella sonnei HH109, two and four copies in two different natural isolates of Escherichia coli, and two copies in E. coli K-12 strain JM109 from the 60 bacterial strains that were examined. All dam sites in the IS10s in E. cloacae MD36 and S. sonnei HH109 were methylated. Tn10 and IS10 transpose by a mechanism in which the element is excised from the donor site and inserted into the new target site without significant replication of the transposing segment; thus, the copy numbers of the elements in the cell are thought to be unchanged in most circumstances. Accumulation of IS10 copies in E. cloacae MD36 has interesting evolutionary implications.

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Selected References

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

  1. Bender J., Kleckner N. Genetic evidence that Tn10 transposes by a nonreplicative mechanism. Cell. 1986 Jun 20;45(6):801–815. doi: 10.1016/0092-8674(86)90555-6. [DOI] [PubMed] [Google Scholar]
  2. Chao L., Vargas C., Spear B. B., Cox E. C. Transposable elements as mutator genes in evolution. Nature. 1983 Jun 16;303(5918):633–635. doi: 10.1038/303633a0. [DOI] [PubMed] [Google Scholar]
  3. Clerget M., Chandler M., Caro L. The structure of R1drd19: a revised physical map of the plasmid. Mol Gen Genet. 1981;181(2):183–191. doi: 10.1007/BF00268425. [DOI] [PubMed] [Google Scholar]
  4. Davis M. A., Simons R. W., Kleckner N. Tn10 protects itself at two levels from fortuitous activation by external promoters. Cell. 1985 Nov;43(1):379–387. doi: 10.1016/0092-8674(85)90043-1. [DOI] [PubMed] [Google Scholar]
  5. Foster T. J., Davis M. A., Roberts D. E., Takeshita K., Kleckner N. Genetic organization of transposon Tn10. Cell. 1981 Jan;23(1):201–213. doi: 10.1016/0092-8674(81)90285-3. [DOI] [PubMed] [Google Scholar]
  6. Green L., Miller R. D., Dykhuizen D. E., Hartl D. L. Distribution of DNA insertion element IS5 in natural isolates of Escherichia coli. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4500–4504. doi: 10.1073/pnas.81.14.4500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hall B. G., Parker L. L., Betts P. W., DuBose R. F., Sawyer S. A., Hartl D. L. IS103, a new insertion element in Escherichia coli: characterization and distribution in natural populations. Genetics. 1989 Mar;121(3):423–431. doi: 10.1093/genetics/121.3.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Halling S. M., Kleckner N. A symmetrical six-base-pair target site sequence determines Tn10 insertion specificity. Cell. 1982 Jan;28(1):155–163. doi: 10.1016/0092-8674(82)90385-3. [DOI] [PubMed] [Google Scholar]
  9. Halling S. M., Simons R. W., Way J. C., Walsh R. B., Kleckner N. DNA sequence organization of IS10-right of Tn10 and comparison with IS10-left. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2608–2612. doi: 10.1073/pnas.79.8.2608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hillen W., Schollmeier K. Nucleotide sequence of the Tn10 encoded tetracycline resistance gene. Nucleic Acids Res. 1983 Jan 25;11(2):525–539. doi: 10.1093/nar/11.2.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kleckner N., Chan R. K., Tye B. K., Botstein D. Mutagenesis by insertion of a drug-resistance element carrying an inverted repetition. J Mol Biol. 1975 Oct 5;97(4):561–575. doi: 10.1016/s0022-2836(75)80059-3. [DOI] [PubMed] [Google Scholar]
  12. Loh S. M., Cram D. S., Skurray R. A. Nucleotide sequence and transcriptional analysis of a third function (Flm) involved in F-plasmid maintenance. Gene. 1988 Jun 30;66(2):259–268. doi: 10.1016/0378-1119(88)90362-9. [DOI] [PubMed] [Google Scholar]
  13. Machida Y., Machida C., Ohtsubo E. A novel type of transposon generated by insertion element IS102 present in a pSC101 derivative. Cell. 1982 Aug;30(1):29–36. doi: 10.1016/0092-8674(82)90008-3. [DOI] [PubMed] [Google Scholar]
  14. Matsutani S., Ohtsubo E. Complete sequence of IS629. Nucleic Acids Res. 1990 Apr 11;18(7):1899–1899. doi: 10.1093/nar/18.7.1899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Mollet B., Iida S., Shepherd J., Arber W. Nucleotide sequence of IS26, a new prokaryotic mobile genetic element. Nucleic Acids Res. 1983 Sep 24;11(18):6319–6330. doi: 10.1093/nar/11.18.6319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Navas J., García-Lobo J. M., León J., Ortíz J. M. Structural and functional analyses of the fosfomycin resistance transposon Tn2921. J Bacteriol. 1985 Jun;162(3):1061–1067. doi: 10.1128/jb.162.3.1061-1067.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nyman K., Nakamura K., Ohtsubo H., Ohtsubo E. Distribution of the insertion sequence IS1 in gram-negative bacteria. Nature. 1981 Feb 12;289(5798):609–612. doi: 10.1038/289609a0. [DOI] [PubMed] [Google Scholar]
  19. Okamoto S., Suzuki Y., Mise K., Nakaya R. Occurrence of chloramphenicol-acetylating enzymes in various gram-negative bacilli. J Bacteriol. 1967 Nov;94(5):1616–1622. doi: 10.1128/jb.94.5.1616-1622.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Orskov I., Orskov F., Jann B., Jann K. Serology, chemistry, and genetics of O and K antigens of Escherichia coli. Bacteriol Rev. 1977 Sep;41(3):667–710. doi: 10.1128/br.41.3.667-710.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Raleigh E. A., Kleckner N. Multiple IS10 rearrangements in Escherichia coli. J Mol Biol. 1984 Mar 15;173(4):437–461. doi: 10.1016/0022-2836(84)90390-5. [DOI] [PubMed] [Google Scholar]
  22. Roberts D., Hoopes B. C., McClure W. R., Kleckner N. IS10 transposition is regulated by DNA adenine methylation. Cell. 1985 Nov;43(1):117–130. doi: 10.1016/0092-8674(85)90017-0. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Sawyer S. A., Dykhuizen D. E., DuBose R. F., Green L., Mutangadura-Mhlanga T., Wolczyk D. F., Hartl D. L. Distribution and abundance of insertion sequences among natural isolates of Escherichia coli. Genetics. 1987 Jan;115(1):51–63. doi: 10.1093/genetics/115.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schollmeier K., Hillen W. Transposon Tn10 contains two structural genes with opposite polarity between tetA and IS10R. J Bacteriol. 1984 Nov;160(2):499–503. doi: 10.1128/jb.160.2.499-503.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  27. Shen M. M., Raleigh E. A., Kleckner N. Physical analysis of Tn10- and IS10-promoted transpositions and rearrangements. Genetics. 1987 Jul;116(3):359–369. doi: 10.1093/genetics/116.3.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Simons R. W., Kleckner N. Translational control of IS10 transposition. Cell. 1983 Sep;34(2):683–691. doi: 10.1016/0092-8674(83)90401-4. [DOI] [PubMed] [Google Scholar]
  29. Smith D. W., Garland A. M., Herman G., Enns R. E., Baker T. A., Zyskind J. W. Importance of state of methylation of oriC GATC sites in initiation of DNA replication in Escherichia coli. EMBO J. 1985 May;4(5):1319–1326. doi: 10.1002/j.1460-2075.1985.tb03779.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  31. Tenzen T., Matsutani S., Ohtsubo E. Site-specific transposition of insertion sequence IS630. J Bacteriol. 1990 Jul;172(7):3830–3836. doi: 10.1128/jb.172.7.3830-3836.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. doi: 10.1016/0378-1119(84)90012-x. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Yoshioka Y., Ohtsubo H., Ohtsubo E. Repressor gene finO in plasmids R100 and F: constitutive transfer of plasmid F is caused by insertion of IS3 into F finO. J Bacteriol. 1987 Feb;169(2):619–623. doi: 10.1128/jb.169.2.619-623.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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