The transposable element En/Spm-encoded TNPA protein contains a DNA binding and a dimerization domain

Trentmann, Stefan; Saedler, Heinz; Gierl, Alfons

doi:10.1007/bf00279548

Cited by 40 publications

(26 citation statements)

References 30 publications

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“…the strong homology in the TIRs, the 3 bp target site duplication, the structural similarities to Spm and Tam1, and homology within the large ORFs contained within introns. For Spm, the sub-terminal repetitive regions have been demonstrated to be important for transposition (reviewed by Gierl, 1996), and the sub-terminal region, consisting of direct and inverted repeats of a 12 bp sequence, is bound by the TNPA protein Trentmann et al, 1993). While our study does not address the function of domains required for Psl transposition, it is noteworthy that Psl has sub-terminal repeats potentially analogous to those of Spm, although of a different nucleotide sequence.…”

Section: Discussionmentioning

confidence: 94%

Psl: a novel Spm‐like transposable element from Petunia hybrida

Snowden

Napoli

1998

The Plant Journal

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Section: Discussionmentioning

confidence: 94%

Psl: a novel Spm‐like transposable element from Petunia hybrida

Snowden

Napoli

1998

The Plant Journal

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“…The maize Ac transposase binds specifically and co-operatively to repetitive ACG and TCG trinucleotides, which are found in more than twenty copies in both 5′ and 3′ subterminal regions, although the Ac transposase also weakly interacts with the terminal repeats (Kunze & Starlinger, 1989;Becker & Kunze, 1997). The TNPA transposase of the En/Spm element binds a 12 bp sequence found in multiple copies within the 5′ and 3′ 300 bp subterminal repeat regions (Gierl et al, 1988;Trentmann et al, 1993). The Arabidopsis transposon Tag1 also requires minimal subterminal sequences and a minimal internal spacer between 238 bp and 325 bp for efficient transposition (Liu et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

piggyBac internal sequences are necessary for efficient transformation of target genomes

Harrell

Handler

et al. 2005

Insect Molecular Biology

101

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A previously reported piggyBac minimal sequence cartridge, which is capable of efficient transposition in embryo interplasmid transposition assays, failed to produce transformants at a significant frequency in Drosophila melanogaster compared with full-length or less extensive internal deletion constructs. We have re-examined the importance of these internal domain (ID) sequences for germline transformation using a PCR strategy that effectively adds increasing lengths of ID sequences to each terminus. A series of these piggyBac ID synthetic deletion plasmids containing the 3xP3-ECFP marker gene are compared for germline transformation of D. melanogaster. Our analyses identify a minimal sequence configuration that is sufficient for movement of piggyBac vectored sequences from plasmids into the insect genome. Southern hybridizations confirm the presence of the piggyBac transposon sequences, and insertion site analyses confirm these integrations target TTAA sites. The results verify that ID sequences adjacent to the 5' and 3' terminal repeat domains are crucial for effective germline transformation with piggyBac even though they are not required for excision or interplasmid transposition. Using this information we reconstructed an inverted repeat cartridge, ITR1.1k, and a minimal piggyBac transposon vector, pXL-BacII-ECFP, each of which contains these identified ID sequences in addition to the terminal repeat configuration previously described as essential for mobility. We confirm in independent experiments that these new minimal constructs yield transformation frequencies similar to the control piggyBac vector. Sequencing analyses of our constructs verify the position and the source of a point mutation within the 3' internal repeat sequence of our vectors that has no apparent effect on transformation efficiency.

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“…Generally, integral CACTA elements code at least for two proteins with distinct functions that are produced from a single precursor transcript by alternative splicing (Pereira et al, 1986;Masson et al, 1989). One protein, named TNPD, represents the putative transposase required for the excision/integration process during transposition; the other one, named TNPA, is a factor with multiple functions, some of them reflecting its ability to bind DNA (Gierl et al, 1989;Trentmann et al, 1993). When TNPA protein is bound to the subterminal domains of defective CACTA elements the RNA polymerase advancement is stopped, resulting in prematurely terminated transcripts.…”

Section: Resultsmentioning

confidence: 99%

Transposon‐dependent induction of Vincent van Gogh's sunflowers: Exceptions revealed

Fambrini

Salvini

Basile

et al. 2014

Genesis

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The radiate sunflower inflorescence is composed by zygomorphic ray flowers and actinomorphic disk flowers. Studies performed on mutants identify HaCYC2c, a CYCLOIDEA (CYC)-like gene, as one of the key players controlling flower symmetry in sunflower. turf and tub mutants are characterized by a shift from zygomorphic to actinomorphic ray flowers, caused by insertion of transposable elements (TEs) in HaCYC2c gene. In dbl or Chry mutants, an insertion upstream the coding region of HaCYC2c causes the ectopic expression of the gene and the shift from actinomorphic to zygomorphic disk flowers. We focused on Chry2 mutant: a 1034 bp insertion placed 558 bp before the start codon of HaCYC2c was identified. The insertion is a truncated version of a CACTA TE. Unexpectedly, phenotypic and genetic co-segregation analysis in F2 and F3 progenies derived from the crosses Chry2 × turf and turf × Chry2 demonstrated that CACTA insertion is not always sufficient to alter the expression of HaCYC2c gene and generate Chry2 phenotype. F3 plants homozygous for the CACTA insertion displayed either HaCYC2c transcription pattern identical to wild-type plants or a normal heterogamous inflorescence. Stated these results, we conclude that a much more complex regulatory system stays behind the Chry2 phenotype.

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The transposable element En/Spm-encoded TNPA protein contains a DNA binding and a dimerization domain

Cited by 40 publications

References 30 publications

Psl: a novel Spm‐like transposable element from Petunia hybrida

Psl: a novel Spm‐like transposable element from Petunia hybrida

piggyBac internal sequences are necessary for efficient transformation of target genomes

Transposon‐dependent induction of Vincent van Gogh's sunflowers: Exceptions revealed

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