Resetting the Site: Redirecting Integration of an Insertion Sequence in a Predictable Way

• Published in: Molecular cell 2009-06, Vol.34 (5), p.612-619
• Main Authors: Guynet, Catherine, Achard, Adeline, Hoang, Bao Ton, Barabas, Orsolya, Hickman, Alison Burgess, Dyda, Frederick, Chandler, Michael
• Format: Article
• Language: English
• Subjects: Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - physiology; Base Pairing; Base Sequence; DNA; DNA Transposable Elements - physiology; DNA, Single-Stranded - chemistry; Helicobacter pylori - genetics; Models, Genetic; Point Mutation; Transposases - chemistry; Transposases - genetics; Transposases - physiology
• ISSN: 1097-2765; 1097-4164
• DOI: 10.1016/j.molcel.2009.05.017

Target site choice is a complex and poorly understood aspect of DNA transposition despite its importance in rational transposon-mediated gene delivery. Though most transposons choose target sites essentially randomly or with some slight sequence or structural preferences, insertion sequence IS608 from Helicobacter pylori, which transposes using single-stranded DNA, always inserts just 3′ of a TTAC tetranucleotide. Our results from studies on the IS608 transposition mechanism demonstrated that the transposase recognizes its target site by co-opting an internal segment of transposon DNA and utilizes it for specific recognition of the target sites through base-pairing. This suggested a way to redirect IS608 transposition to novel target sites. As we demonstrate here, we can now direct insertions in a predictable way into a variety of different chosen target sequences, both in vitro and in vivo.