EMD-4410
Yeast RPA bound to ssDNA
EMD-4410
Single-particle4.7 Å
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Map released: 19/12/2018
Last modified: 15/05/2024
Sample Organism:
Saccharomyces cerevisiae (strain ATCC 204508 / S288c),
synthetic construct
Sample: Replication Protein A heterotrimeric complex with ssDNA
Fitted models: 6i52 (Avg. Q-score: 0.101)
Deposition Authors: Yates LA
,
Aramayo RJ
Sample: Replication Protein A heterotrimeric complex with ssDNA
Fitted models: 6i52 (Avg. Q-score: 0.101)
Deposition Authors: Yates LA
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A structural and dynamic model for the assembly of Replication Protein A on single-stranded DNA.
Yates LA
,
Aramayo RJ,
Pokhrel N,
Caldwell CC
,
Kaplan JA,
Perera RL,
Spies M
,
Antony E
,
Zhang X
(2018) Nat Commun , 9 , 5447 - 5447
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(2018) Nat Commun , 9 , 5447 - 5447
Abstract:
Replication Protein A (RPA), the major eukaryotic single stranded DNA-binding protein, binds to exposed ssDNA to protect it from nucleases, participates in a myriad of nucleic acid transactions and coordinates the recruitment of other important players. RPA is a heterotrimer and coats long stretches of single-stranded DNA (ssDNA). The precise molecular architecture of the RPA subunits and its DNA binding domains (DBDs) during assembly is poorly understood. Using cryo electron microscopy we obtained a 3D reconstruction of the RPA trimerisation core bound with ssDNA (∼55 kDa) at ∼4.7 Å resolution and a dimeric RPA assembly on ssDNA. FRET-based solution studies reveal dynamic rearrangements of DBDs during coordinated RPA binding and this activity is regulated by phosphorylation at S178 in RPA70. We present a structural model on how dynamic DBDs promote the cooperative assembly of multiple RPAs on long ssDNA.
Replication Protein A (RPA), the major eukaryotic single stranded DNA-binding protein, binds to exposed ssDNA to protect it from nucleases, participates in a myriad of nucleic acid transactions and coordinates the recruitment of other important players. RPA is a heterotrimer and coats long stretches of single-stranded DNA (ssDNA). The precise molecular architecture of the RPA subunits and its DNA binding domains (DBDs) during assembly is poorly understood. Using cryo electron microscopy we obtained a 3D reconstruction of the RPA trimerisation core bound with ssDNA (∼55 kDa) at ∼4.7 Å resolution and a dimeric RPA assembly on ssDNA. FRET-based solution studies reveal dynamic rearrangements of DBDs during coordinated RPA binding and this activity is regulated by phosphorylation at S178 in RPA70. We present a structural model on how dynamic DBDs promote the cooperative assembly of multiple RPAs on long ssDNA.