5EAN

Crystal structure of Dna2 in complex with a 5' overhang DNA

Summary for 5EAN

• Entry DOI: 10.2210/pdb5ean/pdb
• Related: 5EAW 5EAX 5EAY
• Descriptor: DNA replication ATP-dependent helicase/nuclease DNA2, DNA (5'-D(P*AP*CP*TP*CP*TP*GP*CP*CP*AP*AP*GP*AP*GP*GP*A)-3'), IRON/SULFUR CLUSTER, ... (6 entities in total)
• Functional Keywords: dna binding protein, hydrolase-dna complex, hydrolase/dna
• Biological source: Mus musculus (Mouse); More
• Cellular location: Nucleus : Q6ZQJ5
• Total number of polymer chains: 2
• Total formula weight: 124642.90

Authors

Zhou, C.,Pourmal, S.,Pavletich, N.P. (deposition date: 2015-10-16, 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 (2.36 Å)