Summary
The Escherichia coli enterotoxin STII gene is carried by Tn4521. The terminal repeats of Tn4521 are composed of IS2 sequences; however, neither repeat is a complete IS2. In order to determine how this seemingly defective transposon could transpose, mutations were generated within Tn4521 to determine the regions essential for transposition. The left terminal repeat region was found to be non-essential, but the right terminal repeat area was demonstrated to be crucial for transposition. Within the right terminal repeat area is an open reading frame (ORF), capable of encoding a 159 amino acid protein, which was shown by frameshift mutation analysis to be required for transposition. This protein may be the transposase of Tn4521. A pair of 11 bp repeat sequences flanking the ORF was also found to be important. The right 11 bp repeat is part of the left IS2 terminal sequence, and the left 11 bp repeat is located about 300 bp upstream from the right IS2 terminal sequence located within the right terminal repeat region. The results of this study suggest that Tn4521 is a functional transposon and that the sequence including this pair of 11 bp sequences plus the intervening sequence is a transposable element which may be responsible for Tn4521 transposition.
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References
Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472
Burgess NM, Bywater PJ, Cowley CM, Mullan NA, Newsome PM (1978) Biological evaluation of a methanol-soluble, heatstable Escherichia coli enterotoxin in infant mice, pigs, rabbits, and calves. Infect Immun 21:526–531
Clements JD, Finkelstein RA (1978) Immunological cross reactivity between a heat-labile enterotoxin(s) of Escherichia coli and subunits of Vibrio cholerae enterotoxin. Infect Immun 21:1036–1039
Davidson N, Deonier RC, Hu S, Ohtsubo E (1974) Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. X. Deoxyribonucleic acid sequence organization of F and F-primes, and the sequences involved in Hfr formation. In: Schlessinger D (ed) Microbiology-1974. American Society for Microbiology, Washington, DC, pp 56–65
Dean AG, Chiang YC, Williams RG, Harden LB (1972) Test for Escherichia coli enterotoxin using infant mice: application in a study of diarrhea in children in Honolulu. J Infect Dis 125:407–411
Gormann CM, Moffat LF, Howard BH (1982) Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol 2:1044–1051
Grindley NDF, Joyce CM (1981) Analysis of the structure and function of the kanamycin-resistance transposon Tn903. Cold Spring Harbor Symp Quant Biol 45:125–133
Guyer MS, Reed RR, Steitz JA, Low KB (1980) Identification of a sex-affinity site in E. coli as gamma-delta. Cold Spring Harbor Symp Quant Biol 44:135–140
Gyles CL (1971) Heat labile and heat stable forms of the enterotoxin from E. coli strains enteropathogenic for gigs. Ann NY Acad Sci 176:314–322
Gyles CL (1974) Immunological study of the heat-labile enterotoxin of Escherichia coli and Vibrio cholerae. Infect Immun 9:564–569
Gyles C, So M, Falkow S (1974) The enterotoxin plasmid of Escherichia coli. J Infect Dis 130:40–49
Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
Hinton DM, Musso RE (1983) Specific in vitro transcription of the insertion sequence IS2. J Mol Biol 169:53–81
Hu ST, Yang MK, Spandau DF, Lee CH (1987) Characterization of the terminal sequences flanking the transposon that carries the Escherichia coli enterotoxin STII gene. Gene 55:157–167
Lee CH, Hu ST, Swiatek PJ, Moseley SL, Allen SD, So M (1985) Isolation of a novel transposon which carries the Escherichia coli enterotoxin STII gene. J Bacteriol 162:615–620
Matthias PD, Bernard HU, Scott A, Brady G, Hashimoto-Gotoh T, Schutz G (1983) A bovine papilloma virus vector with a dominant resistance marker replicates extrachromosomally in mouse and E. coli cells. EMBO J 2:1487–1492
Moon HW, Whipp SC (1970) Development of resistance with age by swine intestine to effects of enteropathogenic Escherichia coli. J Infect Dis 122:220–223
Motsch S, Schmitt R (1984) Replicon fusion mediated by a singlended derivative of transposon Tn1721. Mol Gen Genet 195:281–287
Ronecker H-J, Rak B (1987) Genetic organization of insertion element IS2 based on a revised nucleotide sequence. Gene 59:291–296
Sack RB (1975) Human diarrheal disease caused by enterotoxigenic Escherichia coli. Annu Rev Microbiol 29:333–353
Skerman FJ, Formal SB, Falkow S (1972) Plasmid-associated enterotoxin production in a strain of Escherichia coli isolated from humans. Infect Immun 5:622–624
So M, McCarthy BJ (1980) Nucleotide sequence of the bacterial transposon Tn1681 encoding a heat-stable (ST) toxin and its identification in enterotoxigenic Escherichia coli strains. Proc natl Acad Sci USA 77:4011–4015
So M, Heffron F, McCarthy BJ (1979) The E. coli gene encoding heat stable toxin is a bacterial transposon flanked by inverted repeats of IS1. Nature 277:453–456
Whipp SC, Moon HW, Argenzio RA (1981) Comparison of enterotoxic activities of heat-stable enterotoxins from class 1 and class 2 Escherichia coli of swine origin. Infect Immun 31:245–256
Young R, Grillo DS (1980) Transposition of the kanamycin-resistance transposon Tn903. Mol Gen Genet 178:681–689
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Hu, S.T., Lee, C.H. Characterization of the transposon carrying the STII gene of enterotoxigenic Escherichia coli . Mol Gen Genet 214, 490–495 (1988). https://doi.org/10.1007/BF00330485
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DOI: https://doi.org/10.1007/BF00330485