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 the 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 the solution structure of the RED subdomain and characterize its interactions with the transposon DNA.
[NMR paper] NMR solution structure of the RED subdomain of the Sleeping Beauty transposase.
NMR solution structure of the RED subdomain of the Sleeping Beauty transposase.
Related Articles NMR solution structure of the RED subdomain of the Sleeping Beauty transposase.
Protein Sci. 2017 Mar 27;:
Authors: Konnova TA, Singer CM, Nesmelova IV
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...
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03-28-2017 03:06 PM
NMR solution structure of the RED subdomain of the Sleeping Beauty transposase
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....
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03-27-2017 12:33 PM
[NMR paper] NMR structure of the HIV-1 reverse transcriptase thumb subdomain.
NMR structure of the HIV-1 reverse transcriptase thumb subdomain.
Related Articles NMR structure of the HIV-1 reverse transcriptase thumb subdomain.
J Biomol NMR. 2016 Nov 17;
Authors: Sharaf NG, Brereton AE, Byeon IL, Andrew Karplus P, Gronenborn AM
Abstract
The solution NMR structure of the isolated thumb subdomain of HIV-1 reverse transcriptase (RT)*has been determined. A detailed comparison of the current structure with dozens of the highest resolution crystal structures of this domain in the context of the full-length enzyme...
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11-20-2016 09:20 PM
NMR structure of the HIV-1 reverse transcriptase thumb subdomain
NMR structure of the HIV-1 reverse transcriptase thumb subdomain
Abstract
The solution NMR structure of the isolated thumb subdomain of HIV-1 reverse transcriptase (RT)Â*has been determined. A detailed comparison of the current structure with dozens of the highest resolution crystal structures of this domain in the context of the full-length enzyme reveals that the overall structures are very similar, with only two regions exhibiting local conformational differences. The C-terminal capping pattern of the αH helix is subtly different, and the loop...
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11-19-2016 08:35 PM
[NMR paper] NMR structural analysis of Sleeping Beauty transposase binding to DNA.
NMR structural analysis of Sleeping Beauty transposase binding to DNA.
NMR structural analysis of Sleeping Beauty transposase binding to DNA.
Protein Sci. 2013 Oct 30;
Authors: E Carpentier C, Schreifels JM, Aronovich EL, Carlson DF, Hackett PB, Nesmelova IV
Abstract
The Sleeping Beauty (SB) transposon is the most widely used DNA transposon in genetic applications and is the only DNA transposon thus far in clinical trials for human gene therapy. In the absence of atomic level structural information, the development of SB transposon relied...
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11-20-2013 12:42 AM
NMR structural analysis of Sleeping Beauty transposase binding to DNA
NMR structural analysis of Sleeping Beauty transposase binding to DNA
Abstract
The Sleeping Beauty (SB) transposon is the most widely used DNA transposon in genetic applications and is the only DNA transposon thus far in clinical trials for human gene therapy. In the absence of atomic level structural information, the development of SB transposon relied primarily on the biochemical and genetic homology data. While these studies were successful and have yielded hyperactive transposases, structural information is needed to gain a mechanistic understanding of transposase activity and...
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11-18-2013 10:42 PM
NMR structural analysis of Sleeping Beauty transposase binding to DNA
NMR structural analysis of Sleeping Beauty transposase binding to DNA
ABSTRACT
The Sleeping Beauty (SB) transposon is the most widely used DNA transposon in genetic applications and is the only DNA transposon thus far in clinical trials for human gene therapy. In the absence of atomic level structural information, the development of SB transposon relied primarily on the biochemical and genetic homology data. While these studies were successful and have yielded hyperactive transposases, structural information is needed to gain a mechanistic understanding of transposase activity and...
nmrlearner
Journal club
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10-30-2013 10:44 AM
[NMR paper] Refining the overall structure and subdomain orientation of ribosomal protein S4 delt
Refining the overall structure and subdomain orientation of ribosomal protein S4 delta41 with dipolar couplings measured by NMR in uniaxial liquid crystalline phases.
Related Articles Refining the overall structure and subdomain orientation of ribosomal protein S4 delta41 with dipolar couplings measured by NMR in uniaxial liquid crystalline phases.
J Mol Biol. 1999 Sep 17;292(2):375-87
Authors: Markus MA, Gerstner RB, Draper DE, Torchia DA
Prokaryotic protein S4 initiates assembly of the small ribosomal subunit by binding to 16 S rRNA....
NMR solution structure of the RED subdomain of the Sleeping Beauty transposase
Linear Mode
