Formation of the tandem repeat (IS30)2 and its role in IS30-mediated transpositional DNA rearrangements (original) (raw)
Abstract
Plasmids carrying two IS_30_ elements in the same orientation, as in the composite transposon Tn_2706_, are structurally unstable in Escherichia coli. A primary segregation product is formed by site-specific deletion of the sequences carried between the two IS_30_ elements. The resulting covalently closed replicon carries the two IS_30_ elements as tandem repeats separated by only 2 bp. This (IS_30_)2 structure is extremely unstable, but it can nevertheless be isolated on its vector plasmid and, after purification, can be reintroduced into host cells by transformation. Among the descendants of transformants of recA − bacteria, replicated copies of the introduced (IS_30_)2 structure are still present, together with various kinds of segregation products which provide evidence for the efficient generation of DNA rearrangements. Most abundant is the product of another site-specific recombination between two identical ends of the IS_30_ elements involved, which results in the presence of just one intact IS_30_ on the plasmid. Apart from this, and depending on the presence of appropriate targets for IS_30_ transposition, various transposition products of (IS_30_)2 are also seen. Intramolecular reactions lead to DNA inversions and deletions with breakpoints other than IS_30_ ends. In intermolecular reactions inverse transposition occurs at high frequency and one also obtains simple transposition and cointegration. A mutational study revealed the requirement in cis of one intact IS_30_ transposase gene and of both proximal ends of the two IS_30_ elements concerned not only for the formation of (IS_30_)2, but also for its further rearrangement reactions, including the efficient formation of site-specific deletions. A model is proposed, which postulates that (IS_30_)2 intermediates play a key role in IS_30_ transposition pathways in which the formation of (IS_30_)2 may be rate-limiting. Once this structure is formed, it gives rise to a burst of transpositional rearrangements in the subclone carrying (IS_30_)2. Evolutionary implications of these findings are discussed.
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References
- Ahmed A (1986) Evidence for replicative transposition of Tn_5_ and Tn_9_. J Mol Biol 191:75–84
Google Scholar - Benjamin HW, Kleckner N (1989) Intramolecular transposition by Tn_10_. Cell 59:373–383
Google Scholar - Berg DE (1983) Structural requirement for IS_50_-mediated gene transposition. Proc Natl Acad Sci USA 80:792–796
Google Scholar - Berg DE (1989) Transposon Tn_5_. In: Berg DE, Howe MM (eds) Mobile DNA. American Society for Microbiology, Washington DC, pp 185–210
Google Scholar - Birkenbihl RP, Vielmetter W (1989) Complete maps of IS_1_, IS_2_, IS_3_, IS_4_, IS_5_, IS_30_ and IS_150_ locations in Escherichia coli K12. Mol Gen Genet 220:147–153
Google Scholar - Caparon MG, Scott JR (1989) Excision and insertion of the conjugative transposon Tn_916_ involves a novel recombination mechanism. Cell 59:1027–1034
Google Scholar - Caspers P, Dalrymple B, Iida S, Arber W (1984) IS_30_, a new insertion sequence of Escherichia coli K12. Mol Gen Genet 196:68–73
Google Scholar - Chang ACJ, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156
CAS PubMed Google Scholar - Dalrymple B (1987) Novel rearrangements of IS_30_-carrying plasmids leading to the reactivation of gene expression. Mol Gen Genet 207:413–420
Google Scholar - Dalrymple B, Arber W (1985) Promotion of RNA transcription on the insertion element IS_30_ of E. coli K12. EMBO J 4:2687–2693
Google Scholar - Dalrymple B, Arber W (1986) The characterization of terminators of RNA transcription of IS_30_ and an analysis of their role in IS element-mediated polarity. Gene 44:1–10
Google Scholar - Dalrymple B, Caspers P, Arber W (1984) Nucleotide sequence of the procaryotic mobile genetic element IS_30_. EMBO J 3:2145–2149
Google Scholar - Dente L, Cesareni G, Cortese R (1983) pEMBL: a new family of single stranded plasmids. Nucleic Acids Res 11:1645–1655
Google Scholar - Galas DJ, Chandler M (1989) Bacterial insertion sequences. In: Berg DE, Howe MM (eds) Mobile DNA. American Society for Microbiology, Washington DC, pp 109–162
Google Scholar - Haniford DB, Benjamin HW, Kleckner N (1991) Kinetic and structural analysis of a cleaved donor intermediate and a strand transfer intermediate in Tn_10_ transposition. Cell 64:171–179
Google Scholar - Hirschel BJ, Galas DJ, Chandler M (1982) Cointegrate formation by Tn_5_, but not transposition, is dependent on recA. Proc Natl Acad Sci USA 79:4530–4534
Google Scholar - Lichens-Park A, Syvanen M (1988) Cointegrate formation by IS_50_ requires multiple donor molecules. Mol Gen Genet 211:244–251
Google Scholar - Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Google Scholar - Messing J (1979) A multipurpose cloning system based on a single-stranded DNA bacteriophage M13. Recombinant Tech Bull 2:43–48
Google Scholar - Miller J (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Google Scholar - Morisato D, Kleckner N (1984) Transposase promotes double strand breaks and single strand joints at Tn_10_ termini. Cell 39:181–190
Google Scholar - Morisato D, Kleckner N (1987) Tn_10_ transposition and circle formation in vitro. Cell 51:101–111
Google Scholar - Muster CJ, Shapiro JA (1980) Recombination involving transposable elements: On replicon fusion. Cold Spring Harbor Symp Quant Biol 45:239–242
Google Scholar - Reimmann C, Haas D (1987) Mode of replicon fusion mediated by the duplicated insertion sequence IS_21_ in Escherichia coli. Genetics 115:619–625
Google Scholar - Reimmann C, Hass D (1990) The intA gene of insertion sequence IS_21_ is essential for cleavage at the inner 3′ ends of randomly repeated IS_21_ elements in vitro. EMBO J 9:4055–4063
Google Scholar - Reimmann C, Moore R, Little S, Savioz A, Willetts NS, Haas D (1989) Genetic structure, function and regulation of the transposable element IS_21_. Mol Gen Genet 215:416–424
Google Scholar - Rose AM, Snutch TP (1984) Isolation of the closed circular form of the transposable element Tc_1_ in Caenorhabditis elegans. Nature 311:485–486
Google Scholar - Ruan K, Emmons W (1984) Extrachromosomal copies of transposon Tc_1_ in the nematode Caenorhabditis elegans. Proc Natl Acad Sci USA 81:4018–4022
Google Scholar - Russel M, Kidd S, Kelley MR (1986) An improved filamentous helper phage for generating single-stranded plasmid DNA. Gene 45:333–338
Google Scholar - Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Google Scholar - Scott JN, Kirchman PA, Caparon MG (1988) An intermediate in transposition of the conjugative transposon Tn_916_. Proc Natl Acad Sci USA 85:4809–4813
Google Scholar - Spielmann-Ryser J, Moser M, Kast P, Weber H (1991) Factors determining the frequency of plasmid cointegrate formation mediated by insertion sequence IS_3_ from Escherichia coli. Mol Gen Genet 226:441–448
Google Scholar - Stalder R, Arber W (1989) Characterization of in vitro constructed IS_30_-flanked transposons. Gene 76:187–193
Google Scholar - Stalder R, Caspers P, Olasz F, Arber W (1990) The N-terminal domain of the IS_30_ transposase is able to interact specifically with the terminal inverted repeats of the element. J Biol Chem 265:3757–3762
Google Scholar - Sundaresan V, Freeling M (1987) An extrachromosomal form of the Mu transposons of maize. Proc Natl Acad Sci USA 84:4924–4928
Google Scholar - Umeda M, Ohtsubo E (1990) Mapping of insertion element IS_30_ in the Escherichia coli K12 chromosome. Mol Gen Genet 222:317–322
Google Scholar - Vieira J, Messing J (1982) The pUC plasmids, and M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
Google Scholar - Weinert TA, Derbyshire KM, Hughson FM, Grindley ND (1984) Replicative and conservative transpositional recombination of insertion sequences. Cold Spring Harbor Symp Quant Biol 49:251–260
Google Scholar - Weinert TA, Schaus NA, Grindley ND (1983) Insertion sequence duplication in transpositional recombination. Science 222:755–765
Google Scholar - Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
Google Scholar
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- Institute of Molecular Genetics, Agricultural Biotechnological Center, Szent-Györgyi A. u. 4, H-2101, Gödöllö, Hungary
Ferenc Olasz - Department of Microbiology, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056, Basel, Switzerland
Ferenc Olasz, Rolf Stalder & Werner Arber
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- Ferenc Olasz
You can also search for this author inPubMed Google Scholar - Rolf Stalder
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Communicated by H. Hennecke
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Olasz, F., Stalder, R. & Arber, W. Formation of the tandem repeat (IS_30_)2 and its role in IS_30_-mediated transpositional DNA rearrangements.Molec. Gen. Genet. 239, 177–187 (1993). https://doi.org/10.1007/BF00281616
- Received: 19 October 1992
- Accepted: 29 November 1992
- Issue Date: May 1993
- DOI: https://doi.org/10.1007/BF00281616