Engineered Hyperactive Integrase for Concerted HIV-1 DNA Integration

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Engineered Hyperactive Integrase for Concerted HIV-1 DNA Integration

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Title: Engineered Hyperactive Integrase for Concerted HIV-1 DNA Integration
Author: Li, Min; Jurado, Kellie A.; Lin, Shiqiang; Engelman, Alan; Craigie, Robert

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Citation: Li, Min, Kellie A. Jurado, Shiqiang Lin, Alan Engelman, and Robert Craigie. 2014. “Engineered Hyperactive Integrase for Concerted HIV-1 DNA Integration.” PLoS ONE 9 (8): e105078. doi:10.1371/journal.pone.0105078. http://dx.doi.org/10.1371/journal.pone.0105078.
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Abstract: The DNA cutting and joining reactions of HIV-1 integration are catalyzed by integrase (IN), a viral protein that functions as a tetramer bridging the two viral DNA ends (intasome). Two major obstacles for biochemical and structural studies of HIV-1 intasomes are 1) the low efficiency of assembly with oligonucleotide DNA substrates, and 2) the non-specific aggregation of both intasomes and free IN in the reaction mixture. By fusing IN with a small non-specific DNA binding protein, Sulfolobus solfataricus chromosomal protein Sso7d (PDB: 1BNZ), we have engineered a highly soluble and hyperactive IN. Unlike wild-type IN, it efficiently catalyzes intasome assembly and concerted integration with oligonucleotide DNA substrates. The fusion IN protein also functions to integrate viral reverse transcripts during HIV-infection. The hyperactive HIV-1 IN may assist in facilitating future biochemical and structural studies of HIV-1 intasomes. Understanding the mechanistic basis of the Sso7d-IN fusion protein could provide insight into the factors that have hindered biophysical studies of wild-type HIV-1 IN and intasomes.
Published Version: doi:10.1371/journal.pone.0105078
Other Sources: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4132020/pdf/
Terms of Use: This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAA
Citable link to this page: http://nrs.harvard.edu/urn-3:HUL.InstRepos:12785788
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