2N3F

Solution structure of both dsRBDs of DRB4 along with linker (viz. DRB4(1-153))

2N3F の概要

• エントリーDOI: 10.2210/pdb2n3f/pdb
• 関連するPDBエントリー: 2N3G 2N3H
• NMR情報: BMRB: 25138
• 分子名称: Double-stranded RNA-binding protein 4 (1 entity in total)
• 機能のキーワード: rnai, rna binding protein
• 由来する生物種: Arabidopsis thaliana (mouse-ear cress,thale-cress)
• 細胞内の位置: Nucleus : Q8H1D4
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 16618.72

構造登録者

Deshmukh, M.,Chiliveri, S. (登録日: 2015-05-29, 公開日: 2016-09-07, 最終更新日: 2024-05-01)

主引用文献

Chiliveri, S.C.,Aute, R.,Rai, U.,Deshmukh, M.V.
DRB4 dsRBD1 drives dsRNA recognition in Arabidopsis thaliana tasi/siRNA pathway.
Nucleic Acids Res., 45:8551-8563, 2017

PubMed Abstract: In Arabidopsis thaliana, endogenous trans-acting and exogenous siRNA pathways are initiated by the interaction of DRB4 with trigger dsRNA. Further, DCL4:DRB4 complex cleaves the dsRNA into 21 bp siRNA. Understanding molecular determinants and mechanistic details of dsRNA recognition by DRB4 is vital for inducing long-term RNAi-mediated gene regulation in plants. Here, we present solution structures of individual and concatenated DRB4 dsRBDs and demonstrate modes of dsRNA binding by employing NMR, ITC and site-specific mutagenesis. While both dsRBDs adopt the canonical α-β-β-β-α fold, key structural differences and ms-μs dynamics located at the RNA binding region were observed for dsRBD1. These features favor dsRBD1 to orient itself and make stronger tripartite contact with dsRNA, a feature missing in dsRBD2. Additionally, the inter-domain orientation induced by the linker restricts the mobility of dsRBD2, resulting in the steric hindrance of α1 helix in dsRBD2, and leads in further reduction of its dsRNA binding activity. Our study deciphers functional roles of DRB4 domains by showing that dsRBD1 drives the tasiRNA/siRNA pathway. Furthermore, we identify a potential role of the C-terminal region of DRB4 in protein:protein interaction as it possesses six PxxP motifs, binds to Zn2+ and contains a small structural domain.

PubMed: 28575480
DOI: 10.1093/nar/gkx481

実験手法

SOLUTION NMR