7UB2

Structure of RecT protein from Listeria innoccua phage A118 in complex with 83-mer annealed duplex

Summary for 7UB2

• Entry DOI: 10.2210/pdb7ub2/pdb
• EMDB information: 26434
• Descriptor: RecT, DNA (49-mer) (3 entities in total)
• Functional Keywords: dna recombination, dna annealing, dna binding protein, dna binding protein-dna complex, dna binding protein/dna
• Biological source: Listeria innocua Clip11262; More
• Total number of polymer chains: 12
• Total formula weight: 339553.66

Authors

Bell, C.E.,Caldwell, B.J. (deposition date: 2022-03-14, release date: 2022-12-07, Last modification date: 2024-06-12)

Primary citation

Caldwell, B.J.,Norris, A.S.,Karbowski, C.F.,Wiegand, A.M.,Wysocki, V.H.,Bell, C.E.
Structure of a RecT/Red beta family recombinase in complex with a duplex intermediate of DNA annealing.
Nat Commun, 13:7855-7855, 2022

PubMed Abstract: Some bacteriophage encode a recombinase that catalyzes single-stranded DNA annealing (SSA). These proteins are apparently related to RAD52, the primary human SSA protein. The best studied protein, Redβ from bacteriophage λ, binds weakly to ssDNA, not at all to dsDNA, but tightly to a duplex intermediate of annealing formed when two complementary DNA strands are added to the protein sequentially. We used single particle cryo-electron microscopy (cryo-EM) to determine a 3.4 Å structure of a Redβ homolog from a prophage of Listeria innocua in complex with two complementary 83mer oligonucleotides. The structure reveals a helical protein filament bound to a DNA duplex that is highly extended and unwound. Native mass spectrometry confirms that the complex seen by cryo-EM is the predominant species in solution. The protein shares a common core fold with RAD52 and a similar mode of ssDNA-binding. These data provide insights into the mechanism of protein-catalyzed SSA.

PubMed: 36543802
DOI: 10.1038/s41467-022-35572-z

Experimental method

ELECTRON MICROSCOPY (3.4 Å)