2hwt
From Proteopedia
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NMR solution structure of the Master-Rep protein nuclease domain (2-95) from the Faba Bean Necrotic Yellows Virus
Overview
Nanoviruses are a family of plant viruses that possess a genome of, multiple circular single-stranded DNA (ssDNA) components and are, strikingly similar in their replication mode to the plant geminiviruses, and to the circoviruses that infect birds or mammals. These viruses, multiply by rolling circle replication using virus-encoded multifunctional, replication initiator proteins (Rep proteins) that catalyze the initiation, of replication on a double-stranded DNA (dsDNA) intermediate and the, resolution of the ssDNA into circles. Here we report the solution NMR, three-dimensional structure of the endonuclease domain from the master Rep, (M-Rep) protein of faba bean necrotic yellows virus (FBNYV), a, representative of the nanoviruses. The domain comprises amino acids 2-95, (M-Rep2-95), and its global fold is similar to those previously described, for the gemini- and circovirus Rep endonuclease domains, consisting of a, central 5-stranded antiparallel beta-sheet covered on one side by an, alpha-helix and irregular loops and on the other, more open side of the, domain, by an alpha-helix containing the catalytic tyrosine residue (the, catalytic helix). Longer domain constructs extending to amino acids 117, and 124 were also characterized. They contain an additional alpha-helix, are monomeric, and exhibit catalytic activity indistinguishable from that, of M-Rep2-95. The binding site for the catalytic metal was identified by, paramagnetic broadening and maps to residues on the exposed face of the, central beta-sheet. A comparison with the previously determined Rep, endonuclease domain structures of tomato yellow leaf curl Sardinia virus, (TYLCSV), a geminivirus, and that of porcine circovirus type 2 (PCV2) Rep, allows the identification of a positively charged surface that is most, likely involved in dsDNA binding, and reveals common features shared by, all endonuclease domains of nanovirus, geminivirus, and circovirus Rep, proteins.
About this Structure
2HWT is a Single protein structure of sequence from Faba bean necrotic yellows virus. Full crystallographic information is available from OCA.
Reference
Solution structure of the endonuclease domain from the master replication initiator protein of the nanovirus faba bean necrotic yellows virus and comparison with the corresponding geminivirus and circovirus structures., Vega-Rocha S, Gronenborn B, Gronenborn AM, Campos-Olivas R, Biochemistry. 2007 May 29;46(21):6201-12. Epub 2007 May 2. PMID:17472345
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