SASDDT5: Ribonucleoprotein complex of nonstructural protein sigma NS bound...

基本情報

登録情報

データベース: SASBDB / ID: SASDDT5

試料

Ribonucleoprotein complex of nonstructural protein sigma NS bound to 20mer RNA (NS-RNP20)

• Nonstructural protein sigma NS (protein), Avian orthoreovirus
• 20mer RNA (unstructured) (RNA), 20mer

• 機能・相同性: Reovirus non-structural protein sigma NS / Sigma NS protein / single-stranded RNA binding / RNA-dependent RNA polymerase activity / Nonstructural protein sigma NS
• 生物種: Avian orthoreovirus (ウイルス)
• 引用: ジャーナル: Nucleic Acids Res / 年: 2018; タイトル: Stability of local secondary structure determines selectivity of viral RNA chaperones.; 著者: Jack P K Bravo / Alexander Borodavka / Anders Barth / Antonio N Calabrese / Peter Mojzes / Joseph J B Cockburn / Don C Lamb / Roman Tuma / ; 要旨: To maintain genome integrity, segmented double-stranded RNA viruses of the Reoviridae family must accurately select and package a complete set of up to a dozen distinct genomic RNAs. It is thought that the high fidelity segmented genome assembly involves multiple sequence-specific RNA-RNA interactions between single-stranded RNA segment precursors. These are mediated by virus-encoded non-structural proteins with RNA chaperone-like activities, such as rotavirus (RV) NSP2 and avian reovirus σNS. Here, we compared the abilities of NSP2 and σNS to mediate sequence-specific interactions between RV genomic segment precursors. Despite their similar activities, NSP2 successfully promotes inter-segment association, while σNS fails to do so. To understand the mechanisms underlying such selectivity in promoting inter-molecular duplex formation, we compared RNA-binding and helix-unwinding activities of both proteins. We demonstrate that octameric NSP2 binds structured RNAs with high affinity, resulting in efficient intramolecular RNA helix disruption. Hexameric σNS oligomerizes into an octamer that binds two RNAs, yet it exhibits only limited RNA-unwinding activity compared to NSP2. Thus, the formation of intersegment RNA-RNA interactions is governed by both helix-unwinding capacity of the chaperones and stability of RNA structure. We propose that this protein-mediated RNA selection mechanism may underpin the high fidelity assembly of multi-segmented RNA genomes in Reoviridae.

登録者

• Jack Bravo (University of Leeds)

モデル

試料

• 試料: 名称: Ribonucleoprotein complex of nonstructural protein sigma NS bound to 20mer RNA (NS-RNP20); Entity id: 1025 / 1027
• バッファ: 名称: 25 mM HEPES, 150 mM NaCl / pH: 7.5

要素 #1025

タイプ: protein / 記述: Nonstructural protein sigma NS / 分子量: 40.586 / 分子数: 8 / 由来: Avian orthoreovirus / 参照: UniProt: Q9DH28
配列:

MDNTVRVGVS RNTSGAAGQT LFRNFYLLRC NISADGRNAT KAVQSHFPFL SRAVRCLSPL AAHCADRTLR RDNVKQILTR ELPFSSDLIN YAHHVNSSSL TTSQGVEAAR LVAQVYGEQV PFDHIYPTGS ATYCPGAIAN AISRIMAGFV PREGDDFAPS GPIDYLAADL IAYKFVLPYM LDMVDGRPQI VLPSHTVEEM LTNTSLLNSI DASFGIEARS DQRMTRDAAE MSSRSLNELE DHDQRGRMPW KIMLAMMAAQ LKVELDALAD ERTESQANAH VTSFGSRLFN QMSAFVTIDR ELMELALLIK EQGFAMNPGQ IASKWSLIRR SGPTRPLSGA RLEIRNGNWM IREGDQTLLS VSPARMA

要素 #1027

名称: 20mer / タイプ: RNA / 記述: 20mer RNA (unstructured) / 分子量: 6.399 / 分子数: 2
配列:

CUUUUCAAGA CAUGCAACAA

実験情報

• ビーム: 設備名称: Diamond Light Source B21 / 地域: Oxfordshire / 国: UK / 形状: 1 x 5 mm / 線源: X-ray synchrotron / 波長: 0.1 Å / スペクトロメータ・検出器間距離: 3.9 mm
• 検出器: 名称: Pilatus 2M

スキャン

タイトル: Ribonucleoprotein complex of nonstructural protein sigma NS bound to 20mer RNA (NS-RNP20)
測定日: 2017年2月25日 / 単位: 1/A /

	Min	Max
Q	0.0037	0.37

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 1040 /

	Min	Max
Q	0.0059053	0.235036
P(R) point	1	1040
R	0	380

結果

カーブのタイプ: merged
コメント: SigmaNS ribonucleoprotein complex with 20mer RNA. Sample concentration, UNKNOWN; Sample temperature, UNKNOWN; Exposure time; UNKNOWN.

	実験値	Porod
分子量	323 kDa	-
体積	-	964 nm3

	P(R)	Guinier	Guinier error
前方散乱 I0	0.3067	0.2991	0.001
慣性半径, Rg	8.65 nm	7.82 nm	0.06

	Min	Max
D	-	38
Guinier point	11	45