Springer Nature Publishing
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.
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Yates, Luke A.; Aramayo, Ricardo J.; Pokhrel, Nilisha; Caldwell, Colleen C.; Kaplan, Joshua A.; Perera, Rajika L.; Spies, Maria; Antony, Edwin; and Zhang, Xiaodong, "A Structural and Dynamic Model for the Assembly of Replication Protein A on Single-Stranded DNA" (2018). Biological Sciences Faculty Research and Publications. 669.
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