4R71

Structure of the Qbeta holoenzyme complex in the P1211 crystal form

4R71 の概要

• エントリーDOI: 10.2210/pdb4r71/pdb
• 分子名称: Elongation factor Ts, Elongation factor Tu, RNA-directed RNA polymerase beta chain, 30S ribosomal protein S1 (3 entities in total)
• 機能のキーワード: ob fold, translation, viral protein-ribosomal protein complex, viral protein/ribosomal protein
• 由来する生物種: Escherichia coli; 詳細
• 細胞内の位置: Cytoplasm: P0CE48 P0AG67
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 320374.03

構造登録者

Gytz, H.,Seweryn, P.,Kutlubaeva, Z.,Chetverin, A.B.,Brodersen, D.E.,Knudsen, C.R. (登録日: 2014-08-26, 公開日: 2015-09-23, 最終更新日: 2024-02-28)

主引用文献

Gytz, H.,Mohr, D.,Seweryn, P.,Yoshimura, Y.,Kutlubaeva, Z.,Dolman, F.,Chelchessa, B.,Chetverin, A.B.,Mulder, F.A.,Brodersen, D.E.,Knudsen, C.R.
Structural basis for RNA-genome recognition during bacteriophage Q beta replication.
Nucleic Acids Res., 43:10893-10906, 2015

PubMed Abstract: Upon infection of Escherichia coli by bacteriophage Qβ, the virus-encoded β-subunit recruits host translation elongation factors EF-Tu and EF-Ts and ribosomal protein S1 to form the Qβ replicase holoenzyme complex, which is responsible for amplifying the Qβ (+)-RNA genome. Here, we use X-ray crystallography, NMR spectroscopy, as well as sequence conservation, surface electrostatic potential and mutational analyses to decipher the roles of the β-subunit and the first two oligonucleotide-oligosaccharide-binding domains of S1 (OB1-2) in the recognition of Qβ (+)-RNA by the Qβ replicase complex. We show how three basic residues of the β subunit form a patch located adjacent to the OB2 domain, and use NMR spectroscopy to demonstrate for the first time that OB2 is able to interact with RNA. Neutralization of the basic residues by mutagenesis results in a loss of both the phage infectivity in vivo and the ability of Qβ replicase to amplify the genomic RNA in vitro. In contrast, replication of smaller replicable RNAs is not affected. Taken together, our data suggest that the β-subunit and protein S1 cooperatively bind the (+)-stranded Qβ genome during replication initiation and provide a foundation for understanding template discrimination during replication initiation.

PubMed: 26578560
DOI: 10.1093/nar/gkv1212

実験手法

X-RAY DIFFRACTION (3.21 Å)