Precise excision of bacterial transposon Tn 5 in yeast

Gordenin, Dmitry A.; Трофимова, М В; Shaburova, O. N.; Pavlov, Youri I.; Chernoff, Yury O.; Chekuolene, Y. V.; Proscyavichus, Y. Y.; Sasnauskas, Kęstutis; Janulaitis, A.

doi:10.1007/bf00339607

Cited by 18 publications

(9 citation statements)

References 21 publications

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“…These results support the conclusion that the increase of reversion frequency in Text mutants is due to an increase of precise excision frequency. Previously (Gordenin et al, 1988) we confirmed the proposal about the precise excision of Tn5-13 based on restriction analysis by DNA sequencing.…”

Section: Selection Of Mutants With Increased Frequency Of Tn5 Excisiosupporting

confidence: 84%

“…The deletion of the BamHI-XhoI fragment, DS(BX), was generated in this work; other deletions have been described by Chernoff et ai. (1984) and Gordenin et al (1988).…”

Section: Strains and Plasmidsmentioning

confidence: 98%

“…In our previous work (Gordenin et al, 1988) we have shown that bacterial transposon Tn5 can be precisely excised in the eukaryotic organism Saccharomyces cerevisiae. In bacteria, a special form of illegitimate recombination between direct repeats of 9 base pair (bp) target sequence is considered to be the mechanism of precise excision of Tn5 and structurally related transposon Tn10 (Egner and Berg, 1981;Foster et al, 1981;Collins et al, 1982).…”

Section: Introductionmentioning

confidence: 99%

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Yeast mutants with increased bacterial transposon Tn5 excision

Gordenin

Proscyavichus²,

Malkova

et al. 1991

Yeast

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Five complementing recessive mutations that exhibit increased bacterial transposon Tn5 precise excision in yeast Saccharomyces cerevisiae were obtained by ethylmethanesulfonate treatment. One of these mutations (tex1) was submitted to extensive genetic analysis. tex1 is a recessive temperature-sensitive mutation resulting in a 20-100-fold increase in Tn5 excision. It also has increased frequencies of ochre mutation reversion, of forward mutation to canavanine resistance, and loss of chromosome III or its right arm. The possible mechanism of tex1 effects is discussed.

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Section: Selection Of Mutants With Increased Frequency Of Tn5 Excisiosupporting

confidence: 84%

“…The deletion of the BamHI-XhoI fragment, DS(BX), was generated in this work; other deletions have been described by Chernoff et ai. (1984) and Gordenin et al (1988).…”

Section: Strains and Plasmidsmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

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Yeast mutants with increased bacterial transposon Tn5 excision

Gordenin

Proscyavichus²,

Malkova

et al. 1991

Yeast

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“…[Previously, we reported precise excision of Tn5-13 based on sequencing a limited number ofLYS2 revertants of a lys2::TnS insertion mutation located on a multicopy YEp plasmid (7).] Among seven independent Lys+ Adp+ isolates examined, all arose from imprecise excision (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…We therefore introduced TnM into the yeast Saccharomyces cerevisiae and examined excision from the L YS2 gene (7). The low excision rate (10-9) was increased 100-fold in tex mutants (8).…”

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confidence: 99%

Transposon Tn5 excision in yeast: influence of DNA polymerases alpha, delta, and epsilon and repair genes.

Gordenin

Malkova

Peterzen

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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Interaction between short repeats may be a source of genomic rearrangements and deletions. We investigated possible interactions between short (9 base pairs) direct repeats in yeast by using our previously described system for analyzing bacterial transposon TnS excision in yeast. Mutations of either POL3 or POLI, the proposed structural genes for polymerases 8 and a, respectively, yield high levels of excision at semipermissive temperatures. poi2 (correspond to polymerase E) and po12 pol3 double mutants do not exhibit enhanced excision. A majority of excision events involve direct repeats and are precise; the remaining imprecise excisions occur within or in the vicinity of the repeats. The three DNA repair pathways identified by radi, rad6 and radl8, radSO and rad52 mutations were examined for their possible role in TnS excision; no enhancement was observed in mutants. However, the pol3-stimulated TnS excision was reduced in rad52 and radSO mutants. This suggests the potential for interaction between the systems for DNA double-strand break/recombi- polymerases 8 and a are responsible for lagging-strand synthesis and that polymerase e is responsible for leading-strand synthesis, we suggest that TnS excision is stimulated under conditions of altered lagging-strand synthesis, possibly due to extended opportunities for single-strand interactions between the inverted insertion sequence 1550 repeats of TnS.Short repeats of DNA sequence are a source of genomic rearrangements, duplications, and deletions. In bacteria, precise excision occurs between short, 9-base-pair (bp) direct repeats of a transposon target sequence. Excision is stimulated by the long, internal, inverted insertion sequences (IS). Molecular and genetic studies suggest that such deletion may result from "illegitimate" recombination between short repeats. The most likely mechanism is the "slippage" of a replicating strand (1-3). The same mechanism was proposed for formation of deletions between short direct repeats (4). TnS and TnOO excision in Escherichia coli is influenced by genes involved in recombination (recB, recC), mismatch correction (dam, mutH, mutL, mutS, uvrD), single-stranded DNA-binding protein (ssb), and the proofreading exonuclease subunit of DNA polymerase III (dnaQ) (5,6). Precise excision of a small, 50-bp palindromic TnJO remnant is increased in polA (DNA polymerase I) mutants (6).Based on results from bacteria, transposon excision may provide a means for studying various DNA metabolic processes, including DNA repair, recombination, and replication. We therefore introduced TnM into the yeast Saccharomyces cerevisiae and examined excision from the L YS2 gene (7). The low excision rate (10-9) was increased 100-fold in tex mutants (8). These and several other yeast DNA repair and metabolism mutants have been examined in order to understand the TnS excision process. Mutations in the structural genes for two of three yeast DNA polymerases (polymerase a and 5, but not polymerase E) greatly enhance the excision frequency. These obse...

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