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Literature summary for 2.7.7.B22 extracted from

  • Jiang, X.Y.; Hou, F.; Shen, X.D.; Du, X.D.; Xu, H.L.; Zou, S.M.
    The N-terminal zinc finger domain of Tgf2 transposase contributes to DNA binding and to transposition activity (2016), Sci. Rep., 6, 27101.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.7.B22

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzyme Tgf2TPase and transposons in Escherichia coli strain Rosetta1 (DE3) Carassius auratus

Protein Variants

Protein Variants Comment Organism
D228N/E648Q site-directed mutagenesis Carassius auratus
additional information contruction of two truncated recombinant Tgf2 transposases with deletions in the N-terminal zinc finger domain, S1- and S2-Tgf2TPase, from goldfish cDNAs. Both truncated Tgf2TPases lost their DNA-binding ability in vitro, specifically at the ends of Tgf2 transposon than native L-Tgf2TPase. Mutant S1- and S2-Tgf2TPases mediate gene transfer in the zebrafish genome in vivo at a significantly lower efficiency (21%-25%), in comparison with L-Tgf2TPase (56% efficiency). Compared to L-Tgf2TPase, truncated Tgf2TPases catalyze imprecise excisions with partial deletion of TE ends and/or plasmid backbone insertion/deletion. The gene integration into the zebrafish genome mediated by truncated Tgf2TPases is imperfect, creating incomplete 8-bp target site duplications at the insertion sites Carassius auratus

Localization

Localization Comment Organism GeneOntology No. Textmining
nucleus a monopartite nuclear localization signal (NLS, 656-670 aa) is located at the C-terminus Carassius auratus 5634
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Organism

Organism UniProt Comment Textmining
Carassius auratus H9AER4
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information Tgf2 is an autonomously active vertebrate transposon that is efficient at gene-transfer in teleost fish. The N-terminal zinc finger domain of Tgf2 transposase contributes to DNA binding and to transposition activity. Proposed model for Tgf2 transposition, overview Carassius auratus ?
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 ?

Subunits

Subunits Comment Organism
More the full length Tgf2 transposase (L-Tgf2TPase) consisted of several functional domains: an N-terminal BED zinc finger domain (Cx2Cx19Hx4H, 65-120 aa) involved in DNA binding, dimerization domain defined by amino acids 153-213, presumably involved in the formation of oligomers, as well as in DNA binding, and a C-terminus RNase-H domain comprises amino acids 211-683 presumably the core catalytic domain for DNA excision and transposition. Three conserved amino acids residues (DDE) are identified in the RNase-H catalytic domain of Tgf2 transposase, residues of the DDE (D228, D295 and E648) are extremely close in their spatial distribution. A CX2H motif within the RNase-H catalytic domain of Tgf2 transposase is also identified, which which functions as insertion domain for the correct positioning of the final E648 residue of the catalytic triad in the active site. A monopartite nuclear localization signal (NLS, 656-670 aa) is found at the C-terminus. Domain organization, modeling, overview Carassius auratus

Synonyms

Synonyms Comment Organism
Tgf2 transposase
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 Carassius auratus
Tgf2TPase
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 Carassius auratus

General Information

General Information Comment Organism
evolution Tgf2 belongs to the hAT superfamily of transposons, Carassius auratus Tgf2 transposon is another autonomously active vertebrate hAT transposon Carassius auratus
malfunction two truncated recombinant Tgf2 transposases with deletions in the N-terminal zinc finger domain, S1- and S2-Tgf2TPase, from goldfish cDNAs both losing their DNA-binding ability in vitro, specifically at the ends of Tgf2 transposon. Mutant S1- and S2-Tgf2TPases mediate gene transfer in the zebrafish genome in vivo at a significantly lower efficiency (21%-25%), in comparison with L-Tgf2TPase (56% efficiency). Compared to L-Tgf2TPase, truncated Tgf2TPases catalyze imprecise excisions with partial deletion of TE ends and/or plasmid backbone insertion/deletion. The gene integration into the zebrafish genome mediated by truncated Tgf2TPases is imperfect, creating incomplete 8-bp target site duplications at the insertion sites. N-terminal truncated Tgf2 transposases lose their DNA-binding activity Carassius auratus
additional information the Tgf2 element is 4,720 bp long, and the full length Tgf2 transposase is 686 aa long, structure modeling, overview Carassius auratus
physiological function Tgf2 is an autonomously active vertebrate transposon that is efficient at gene-transfer in teleost fish Carassius auratus