Crystal structure of the Wheat dwarf virus Rep domain. Issue 12 (4th December 2019)

• Record Type: Journal Article
• Authors: Everett, Blake A.; Litzau, Lauren A.; Tompkins, Kassidy; Shi, Ke; Nelson, Andrew; Aihara, Hideki; Evans, Robert L.; Gordon, Wendy R.
• Is Part Of: Acta crystallographica. Volume 75:Issue 12(2019:Dec.)
• Page Start: 744
• Page End: 749
• Publication Date: 2019-12-04
• Subjects: crystal structure -- Wheat dwarf virus -- Rep domain -- HUH motif -- HUH‐tag -- ssDNA -- engineered protein–ssDNA complexes; Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X19015796
• Languages: English
• ISSNs: 2053-230X

Abstract

Abstract : The solution of the crystal structure of the Rep domain of Wheat dwarf virus can help inform the development of protein–ssDNA binding tags. Abstract : The Rep domain of Wheat dwarf virus (WDV Rep) is an HUH endonuclease involved in rolling‐circle replication. HUH endonucleases coordinate a metal ion to enable the nicking of a specific ssDNA sequence and the subsequent formation of an intermediate phosphotyrosine bond. This covalent protein–ssDNA adduct makes HUH endonucleases attractive fusion tags (HUH‐tags) in a diverse number of biotechnological applications. Solving the structure of an HUH endonuclease in complex with ssDNA will provide critical information about ssDNA recognition and sequence specificity, thus enabling rationally engineered protein–DNA interactions that are programmable. The structure of the WDV Rep domain reported here was solved in the apo state from a crystal diffracting to 1.24 Å resolution and represents an initial step in the direction of solving the structure of a protein–ssDNA complex.