EMDB/PDB/SASBDBから引用されている文献のデータベース

キーワード
構造解析手法	All X線回折 NMR 電子顕微鏡小角散乱中性子線回折線維回折粉末回折赤外分光 EPR FRET 理論モデルハイブリッド
著者・登録者
ジャーナル
IF	>30 >20 >10 >5 all

タイトル	Distinct Conformational States Underlie Pausing during Initiation of HIV-1 Reverse Transcription.
ジャーナル・号・ページ	J Mol Biol, Vol. 432, Issue 16, Page 4499-4522, Year 2020
掲載日	2020年7月24日
著者	Kevin P Larsen / Junhong Choi / Lynnette N Jackson / Kalli Kappel / Jingji Zhang / Betty Ha / Dong-Hua Chen / Elisabetta Viani Puglisi /
PubMed 要旨	A hallmark of the initiation step of HIV-1 reverse transcription, in which viral RNA genome is converted into double-stranded DNA, is that it is slow and non-processive. Biochemical studies have ...A hallmark of the initiation step of HIV-1 reverse transcription, in which viral RNA genome is converted into double-stranded DNA, is that it is slow and non-processive. Biochemical studies have identified specific sites along the viral RNA genomic template in which reverse transcriptase (RT) stalls. These stalling points, which occur after the addition of three and five template dNTPs, may serve as checkpoints to regulate the precise timing of HIV-1 reverse transcription following viral entry. Structural studies of reverse transcriptase initiation complexes (RTICs) have revealed unique conformations that may explain the slow rate of incorporation; however, questions remain about the temporal evolution of the complex and features that contribute to strong pausing during initiation. Here we present cryo-electron microscopy and single-molecule characterization of an RTIC after three rounds of dNTP incorporation (+3), the first major pausing point during reverse transcription initiation. Cryo-electron microscopy structures of a +3 extended RTIC reveal conformational heterogeneity within the RTIC core. Three distinct conformations were identified, two of which adopt unique, likely off-pathway, intermediates in the canonical polymerization cycle. Single-molecule Förster resonance energy transfer experiments confirm that the +3 RTIC is more structurally dynamic than earlier-stage RTICs. These alternative conformations were selectively disrupted through structure-guided point mutations to shift single-molecule Förster resonance energy transfer populations back toward the on-pathway conformation. Our results support the hypothesis that conformational heterogeneity within the HIV-1 RTIC during pausing serves as an additional means of regulating HIV-1 replication.
リンク	J Mol Biol / PubMed:32512005 / PubMed Central
手法	EM (単粒子)
解像度	4.1 - 6.6 Å
構造データ	21582 6waz EMDB-21582, PDB-6waz: +1 extended HIV-1 reverse transcriptase initiation complex core (pre-translocation state) 手法: EM (単粒子) / 解像度: 4.1 Å 21583 6wb0 EMDB-21583, PDB-6wb0: +3 extended HIV-1 reverse transcriptase initiation complex core (pre-translocation state) 手法: EM (単粒子) / 解像度: 4.2 Å 21584 6wb1 EMDB-21584, PDB-6wb1: +3 extended HIV-1 reverse transcriptase initiation complex core (intermediate state) 手法: EM (単粒子) / 解像度: 4.7 Å 21585 6wb2 EMDB-21585, PDB-6wb2: +3 extended HIV-1 reverse transcriptase initiation complex core (displaced state) 手法: EM (単粒子) / 解像度: 4.5 Å EMDB-22002: +1 extended HIV-1 reverse transcriptase initiation complex (pre-translocation state, global map) 手法: EM (単粒子) / 解像度: 6.6 Å EMDB-22003: +3 extended HIV-1 reverse transcriptase initiation complex (pre-translocation state, unmasked map) 手法: EM (単粒子) / 解像度: 4.9 Å EMDB-22004: +3 extended HIV-1 reverse transcriptase initiation complex (intermediate state, unmasked map) 手法: EM (単粒子) / 解像度: 5.3 Å EMDB-22005: +3 extended HIV-1 reverse transcriptase initiation complex (displaced state, unmasked map) 手法: EM (単粒子) / 解像度: 5.3 Å
由来	human immunodeficiency virus 1 (ヒト免疫不全ウイルス) homo sapiens (ヒト) human immunodeficiency virus type 1 group m subtype b (isolate bh10) (ヒト免疫不全ウイルス)
キーワード	VIRAL PROTEIN/RNA / reverse transcriptase / RNA / complex / protein-RNA complex / tRNA / polymerase / VIRAL PROTEIN / VIRAL PROTEIN-RNA complex