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Functional characterization of an active Rag-like transposase

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Abstract

The formation of diverse immunoglobulin genes results in part from Rag protein–mediated DNA double-strand breaks at the edges of immunoglobulin gene segments, followed by combinatorial reassembly of these segments. We report that a Transib transposase from the insect Helicoverpa zea is active in vitro and that its breakage and joining activities mimic those of Rag, providing strong evidence that Rag and Transib transposases were derived from a common progenitor.

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Figure 1: Hztransib transposase can promote coupled cleavage and strand transfer.
Figure 2: Analysis of the Hztransib DNA-cleavage mechanism.
Figure 3: Hztransib transposition in vitro using precleaved transposon ends.

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Acknowledgements

We thank the other members of the Craig lab for fruitful discussions, H. McComas for her assistance with the figures and text, and S. Desiderio for his comments on the manuscript. N.L.C. is supported as a Howard Hughes Medical Institute Investigator.

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Contributions

N.L.C. conceived the project; C.G.H., X.L. and N.L.C. designed the experiments; C.G.H. and X.L. carried out the experiments; C.G.H., X.L. and N.L.C. analyzed the data and wrote the paper.

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Correspondence to Nancy L Craig.

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The authors declare no competing financial interests.

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Hencken, C., Li, X. & Craig, N. Functional characterization of an active Rag-like transposase. Nat Struct Mol Biol 19, 834–836 (2012). https://doi.org/10.1038/nsmb.2338

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