5U91

Crystal structure of Tre/loxLTR complex

Summary for 5U91

• Entry DOI: 10.2210/pdb5u91/pdb
• Descriptor: Tre recombinase protein, DNA (37-MER), ... (4 entities in total)
• Functional Keywords: cre mutant tre recombinase, isomerase-dna complex, isomerase/dna
• Biological source: synthetic construct; More
• Total number of polymer chains: 8
• Total formula weight: 200710.94

Authors

Meinke, G.,Karpinski, J.,Buchholz, F.,Bohm, A. (deposition date: 2016-12-15, release date: 2017-07-26, Last modification date: 2024-11-06)

Primary citation

Meinke, G.,Karpinski, J.,Buchholz, F.,Bohm, A.
Crystal structure of an engineered, HIV-specific recombinase for removal of integrated proviral DNA.
Nucleic Acids Res., 45:9726-9740, 2017

PubMed Abstract: As part of the HIV infection cycle, viral DNA inserts into the genome of host cells such that the integrated DNA encoding the viral proteins is flanked by long terminal repeat (LTR) regions from the retrovirus. In an effort to develop novel genome editing techniques that safely excise HIV provirus from cells, Tre, an engineered version of Cre recombinase, was designed to target a 34-bp sequence within the HIV-1 LTR (loxLTR). The sequence targeted by Tre lacks the symmetry present in loxP, the natural DNA substrate for Cre. We report here the crystal structure of a catalytically inactive (Y324F) mutant of this engineered Tre recombinase in complex with the loxLTR DNA substrate. We also report that 17 of the 19 amino acid changes relative to Cre contribute to the altered specificity, even though many of these residues do not contact the DNA directly. We hypothesize that some mutations increase the flexibility of the Cre tetramer and that this, along with flexibility in the DNA, enable the engineered enzyme and DNA substrate to adopt complementary conformations.

PubMed: 28934476
DOI: 10.1093/nar/gkx603

Experimental method

X-RAY DIFFRACTION (3.104 Å)