Multivalent interactions essential for lentiviral integrase function.
Multivalent interactions essential for lentiviral integrase function.
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慢病毒整合酶功能的多价相互作用。
DOI：
10.1038/s41467-022-29928-8
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发表时间：
2022-05-03
影响因子：
16.6
通讯作者：
Cherepanov, Peter
中科院分区：
文献类型：
--
作者：
Ballandras-Colas, Allison;Chivukula, Vidya;Gruszka, Dominika T.;Shan, Zelin;Singh, Parmit K.;Pye, Valerie E.;McLean, Rebecca K.;Bedwell, Gregory J.;Li, Wen;Nans, Andrea;Cook, Nicola J.;Fadel, Hind J.;Poeschla, Eric M.;Griffiths, David J.;Vargas, Javier;Taylor, Ian A.;Lyumkis, Dmitry;Yardimci, Hasan;Engelman, Alan N.;Cherepanov, Peter
逆转录病毒整合酶（IN）的多聚体将病毒DNA末端连接在一个稳定的侵入体核蛋白复合体内，以便整合到宿主细胞基因组中。对绵羊慢病毒梅迪-维斯纳病毒（MVV）的侵入体进行重组，发现了一个包含16个IN亚基的大组装体1。在此，我们报告了慢病毒切入体在目标DNA接触之前和链转移之后的低温电镜结构，分别以3.4和3.5 Å分辨率精制。这些结构阐明了慢病毒侵入体形成过程中蛋白质-蛋白质和蛋白质- dna界面的细节。我们发现，参与in十六进制形成的同源界面和连接c端和催化核心结构域的α-螺旋结构对于MVV体外in链转移活性和病毒传染性至关重要。结合光漂白的单分子显微镜显示，MVV侵入体可以结合可变数量的慢病毒特异性宿主因子LEDGF/p75，最多可达16个分子。同样，内源性LEDGF/p75的消融导致人与羊细胞中MVV整合位点的总体重新分布。我们的数据证实了在慢病毒内体的冷冻电镜研究中观察到的扩展结构的重要性，并表明这种组织是与染色质进行多价相互作用以整合活性基因的基础。作者确定了慢病毒侵入体（将病毒DNA插入宿主染色体的核蛋白复合体）的高分辨率低温电镜结构，并表明由16个整合酶亚基组成的结构对其功能至关重要。
A multimer of retroviral integrase (IN) synapses viral DNA ends within a stable intasome nucleoprotein complex for integration into a host cell genome. Reconstitution of the intasome from the maedi-visna virus (MVV), an ovine lentivirus, revealed a large assembly containing sixteen IN subunits1. Herein, we report cryo-EM structures of the lentiviral intasome prior to engagement of target DNA and following strand transfer, refined at 3.4 and 3.5 Å resolution, respectively. The structures elucidate details of the protein-protein and protein-DNA interfaces involved in lentiviral intasome formation. We show that the homomeric interfaces involved in IN hexadecamer formation and the α-helical configuration of the linker connecting the C-terminal and catalytic core domains are critical for MVV IN strand transfer activity in vitro and for virus infectivity. Single-molecule microscopy in conjunction with photobleaching reveals that the MVV intasome can bind a variable number, up to sixteen molecules, of the lentivirus-specific host factor LEDGF/p75. Concordantly, ablation of endogenous LEDGF/p75 results in gross redistribution of MVV integration sites in human and ovine cells. Our data confirm the importance of the expanded architecture observed in cryo-EM studies of lentiviral intasomes and suggest that this organization underlies multivalent interactions with chromatin for integration targeting to active genes. The authors determined high-resolution cryo-EM structures of the lentiviral intasome — the nucleoprotein complex that inserts viral DNA into a host chromosome — and show that the architecture comprising 16 integrase subunits is critical for its function.
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影响因子：
14.9
作者：
Bedwell GJ;Engelman AN
通讯作者：
Engelman AN
影响因子：
4.8
作者：
Cherepanov, P;Maertens, G;Debyser, Z
通讯作者：
Debyser, Z
影响因子：
3.3
作者：
Benleulmi MS;Matysiak J;Henriquez DR;Vaillant C;Lesbats P;Calmels C;Naughtin M;Leon O;Skalka AM;Ruff M;Lavigne M;Andreola ML;Parissi V
通讯作者：
Parissi V
影响因子：
3.7
作者：
ANDRESSON, OS;ELSER, JE;PETURSSON, G
通讯作者：
PETURSSON, G
DOI：
10.1073/pnas.150220297
发表时间：
2000-07-18
影响因子：
11.1
作者：
Chen, JCH;Krucinski, J;Stroud, RM
通讯作者：
Stroud, RM