EMD-4410

Single-particle
4.7 Å
EMD-4410 Deposition: 12/11/2018
Map released: 19/12/2018
Last modified: 15/05/2024
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links

EMD-4410

Yeast RPA bound to ssDNA

EMD-4410

Single-particle
4.7 Å
EMD-4410 Deposition: 12/11/2018
Map released: 19/12/2018
Last modified: 15/05/2024
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
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
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
PUBMED: 30575763
DOI: doi:10.1038/s41467-018-07883-7
ISSN: 2041-1723
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.