resurrected mammalian hAT transposable element and a closely related insect element are highly active in human cell culture

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 110; no. 6; pp. E478 - E487
• Main Authors: Li, Xianghong, Ewis, Hosam, Hice, Robert H, Malani, Nirav, Parker, Nicole, Zhou, Liqin, Feschotte, Cédric, Bushman, Frederic D, Atkinson, Peter W, Craig, Nancy L
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 05.02.2013; National Acad Sciences
• Series: PNAS Plus
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Binding Sites - genetics; Biological Sciences; Cell culture; Chromosomes; Deoxyribonucleic acid; DNA; DNA Transposable Elements - genetics; Gene Transfer, Horizontal; Genes, Insect; Genetic Engineering; Genomes; HeLa Cells; Humans; Molecular Sequence Data; Phylogeny; PNAS Plus; Proteins; Saccharomyces cerevisiae - genetics; Sequence Homology, Amino Acid; Species Specificity; Transposases - metabolism; Tribolium - genetics; Yeast
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1121543109

Chromosome structure and function are influenced by transposable elements, which are mobile DNA segments that can move from place to place. hAT elements are a superfamily of DNA cut and paste elements that move by excision and integration. We have characterized two hAT elements, TcBuster and Space Invaders (SPIN), that are members of a recently described subfamily of hAT elements called Buster elements. We show that TcBuster , from the red flour beetle Tribolium castaneum , is highly active in human cells. SPIN elements are currently inactive elements that were recently highly active in multiple vertebrate genomes, and the high level of sequence similarity across widely diverged species and patchy phylogenetic distribution suggest that they may have moved between genomes by horizontal transfer. We have generated an intact version of this element, SPIN ON, which is highly active in human cells. In vitro analysis of TcBuster and SPIN ON shows that no proteins other than transposase are essential for recombination, a property that may contribute to the ability of SPIN to successfully invade multiple organisms. We also analyze the target site preferences of de novo insertions in the human genome of TcBuster and SPIN ON and compare them with the preferences of Sleeping Beauty and piggyBac , showing that each superfamily has a distinctive pattern of insertion. The high-frequency transposition of both TcBuster and SPIN ON suggests that these transposon systems offer powerful tools for genome engineering. Finally, we describe a Saccharomyces cerevisiae assay for TcBuster that will provide a means for isolation of hyperactive and other interesting classes of transposase mutants.