We have been investigating casposons, proposed to be novel mobile elements that may have been the precursor of bacterial and archaeal adaptive immune systems. We first examined the DNA sequences surrounding these proposed mobile elements, and discovered that many of them are enclosed within short, duplicated segments of genomic DNA of 14-15 basepairs. This was new and compelling evidence that a hallmark of DNA transposition - target site duplications - was also associated with casposons. We successfully expressed and purified the proposed transposase associated with one particular casposon, that from a deep sea vent archaeal species, Aciduliprofundum boonei. We have probed the biochemical properties of this casposase, and have shown that the protein can integrate both short oligonucleotides and also a 2.8 kb excised mini-casposon construct into target DNA. Casposon integration occurs without target specificity into pUC19 and generates 14-15 basepair target site duplications, consistent with those found in casposon host genomes. This result serves as the basis for our ongoing studies to structurally characterize casposases bound to their DNA substrates. We have recently widened our efforts to include other well-behaving and soluble casposases from other archaeal species.
| Dyda, Fred; Hickman, Alison B (2015) Mechanism of spacer integration links the CRISPR/Cas system to transposition as a form of mobile DNA. Mob DNA 6:9 |
| Hickman, Alison B; Dyda, Fred (2015) The casposon-encoded Cas1 protein from Aciduliprofundum boonei is a DNA integrase that generates target site duplications. Nucleic Acids Res 43:10576-87 |
