5EAY

Crystal structure of a Dna2 peptide in complex with Rpa 70N

Summary for 5EAY

• Entry DOI: 10.2210/pdb5eay/pdb
• Related: 5EAN 5EAW 5EAX
• Descriptor: Replication protein A 70 kDa DNA-binding subunit, DNA replication ATP-dependent helicase/nuclease DNA2 (2 entities in total)
• Functional Keywords: dna binding protein
• Biological source: Homo sapiens (Human); More
• Cellular location: Nucleus : P27694 P51530
• Total number of polymer chains: 8
• Total formula weight: 58503.72

Authors

Zhou, C.,Pourmal, S.,Pavletich, N.P. (deposition date: 2015-10-17, release date: 2015-11-18, Last modification date: 2023-09-27)

Primary citation

Zhou, C.,Pourmal, S.,Pavletich, N.P.
Dna2 nuclease-helicase structure, mechanism and regulation by Rpa.
Elife, 4:-, 2015

PubMed Abstract: The Dna2 nuclease-helicase maintains genomic integrity by processing DNA double-strand breaks, Okazaki fragments and stalled replication forks. Dna2 requires ssDNA ends, and is dependent on the ssDNA-binding protein Rpa, which controls cleavage polarity. Here we present the 2.3 Å structure of intact mouse Dna2 bound to a 15-nucleotide ssDNA. The nuclease active site is embedded in a long, narrow tunnel through which the DNA has to thread. The helicase domain is required for DNA binding but not threading. We also present the structure of a flexibly-tethered Dna2-Rpa interaction that recruits Dna2 to Rpa-coated DNA. We establish that a second Dna2-Rpa interaction is mutually exclusive with Rpa-DNA interactions and mediates the displacement of Rpa from ssDNA. This interaction occurs at the nuclease tunnel entrance and the 5' end of the Rpa-DNA complex. Hence, it only displaces Rpa from the 5' but not 3' end, explaining how Rpa regulates cleavage polarity.

PubMed: 26491943
DOI: 10.7554/eLife.09832

Experimental method

X-RAY DIFFRACTION (1.55 Å)