EMD-26434
Structure of RecT protein from Listeria innoccua phage A118 in complex with 83-mer annealed duplex
EMD-26434
Single-particle3.4 Å
Deposition: 14/03/2022Map released: 07/12/2022
Last modified: 12/06/2024
Sample Organism:
Listeria innocua Clip11262,
Escherichia virus M13
Sample: RecT protein from Listeria innocua phage A118, complexed with two complementary strands of ssDNA that were added to the protein sequentially
Fitted models: 7ub2 (Avg. Q-score: 0.508)
Raw data: EMPIAR-11348
Deposition Authors: Bell CE
,
Caldwell BJ
Sample: RecT protein from Listeria innocua phage A118, complexed with two complementary strands of ssDNA that were added to the protein sequentially
Fitted models: 7ub2 (Avg. Q-score: 0.508)
Raw data: EMPIAR-11348
Deposition Authors: Bell CE
,
Caldwell BJ
Structure of a RecT/Red beta family recombinase in complex with a duplex intermediate of DNA annealing.
Caldwell BJ,
Norris AS,
Karbowski CF ,
Wiegand AM,
Wysocki VH ,
Bell CE
(2022) Nat Commun , 13 , 7855 - 7855
,
Wiegand AM,
Wysocki VH ,
Bell CE
(2022) Nat Commun , 13 , 7855 - 7855
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.
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.