2IHX

Solution Structure of the Rous Sarcoma Virus Nucleocapsid Protein:uPsi RNA Packaging Signal Complex

2IHX の概要

• エントリーDOI: 10.2210/pdb2ihx/pdb
• NMR情報: BMRB: 15113
• 分子名称: uPsi RNA, Nucleocapsid (NC) Protein, ZINC ION (3 entities in total)
• 機能のキーワード: protein-rna complex, viral protein-rna complex, viral protein/rna
• 由来する生物種: Rous sarcoma virus; 詳細
• 細胞内の位置: Matrix protein p19: Virion (Potential). Capsid protein p27: Virion (Potential). Nucleocapsid protein p12: Virion (Potential): P03322
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 31104.89

構造登録者

Zhou, J.,Summers, M. (登録日: 2006-09-27, 公開日: 2007-01-16, 最終更新日: 2024-05-29)

主引用文献

Zhou, J.,Bean, R.L.,Vogt, V.M.,Summers, M.
Solution Structure of the Rous Sarcoma Virus Nucleocapsid Protein: muPsi RNA Packaging Signal Complex.
J.Mol.Biol., 365:453-467, 2007

PubMed Abstract: The 5'-untranslated region (5'-UTR) of retroviral genomes contains elements required for genome packaging during virus assembly. For many retroviruses, the packaging elements reside in non-contiguous segments that span most or all of the 5'-UTR. The Rous sarcoma virus (RSV) is an exception, in that its genome can be packaged efficiently by a relatively short, 82 nt segment of the 5'-UTR called muPsi. The RSV 5'-UTR also contains three translational start codons (AUG-1, AUG-2 and AUG-3) that have been controvertibly implicated in translation initiation and genome packaging, one of which (AUG-3) resides within the muPsi sequence. We demonstrated recently that muPsi is capable of binding to the cognate RSV nucleocapsid protein (NC) with high affinity (dissociation constant K(d) approximately 2 nM), and that residues of AUG-3 are essential for tight binding. We now report the solution structure of the NC:muPsi complex, determined using NMR data obtained for samples containing ((13)C,(15)N)-labeled NC and (2)H-enriched, nucleotide-specifically protonated RNAs. Upon NC binding, muPsi adopts a stable secondary structure that consists of three stem loops (SL-A, SL-B and SL-C) and an 8 bp stem (O3). Binding is mediated by the two zinc knuckle domains of NC. The N-terminal knuckle interacts with a conserved U(217)GCG tetraloop (a member of the UNCG family; N=A,U,G or C), and the C-terminal zinc knuckle binds to residues that flank SL-A, including residues of AUG-3. Mutations of critical nucleotides in these sequences compromise or abolish viral infectivity. Our studies reveal novel structural features important for NC:RNA binding, and support the hypothesis that AUG-3 is conserved for genome packaging rather than translational control.

PubMed: 17070546
DOI: 10.1016/j.jmb.2006.10.013

実験手法

SOLUTION NMR