NMR solution structure of the RED subdomain of the Sleeping Beauty transposase
Abstract
DNA transposons can be employed for stable gene transfer in vertebrates. The Sleeping Beauty (SB) DNA transposon has been recently adapted for human application and is being evaluated in clinical trials, however its molecular mechanism is not clear. SB transposition is catalyzed by the transposase enzyme, which is a multi-domain protein containing the catalytic and the DNA-binding domains. The DNA-binding domain of SB transposase contains two structurally independent subdomains, PAI and RED. Recently, the structures of the catalytic domain and the PAI subdomain have been determined, however no structural information on the RED subdomain and its interactions with DNA has been available. Here, we used NMR spectroscopy to determine solution structure of RED subdomain and characterize its interactions with the transposon DNA.
Significance statement: The (SB) transposon is the most widely used DNA transposon in genetic applications and is the only DNA transposon adapted for human gene therapy. Therefore, it is particularly important to have a mechanistic understanding of SB transposase activity. Structural information at the atomic level is vital for such understanding. We report the structure of the last missing piece of SB transposase, the RED subdomain and provide insight into its folding and DNA-binding properties. This article is protected by copyright. All rights reserved.
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