National Institutes of Health/National Institute of General Medical Sciences
R01-GM107013
米国
National Institutes of Health/National Institute Of Allergy and Infectious Diseases
R01-AI147890
米国
National Institutes of Health/National Institute of General Medical Sciences
P30-GM110758
米国
National Science Foundation (United States)
1659534
米国
National Institutes of Health/National Institute Of Allergy and Infectious Diseases
AI142263
米国
National Institutes of Health/National Institute Of Allergy and Infectious Diseases
P50AI150481
米国
Medical Research Council (United Kingdom)
MC_UP_1201/16
英国
European Research Council
ERC-2014-CoG 648432 - MEMBRANEFUSION
英国
引用
ジャーナル: PLoS Pathog / 年: 2020 タイトル: Structures of immature EIAV Gag lattices reveal a conserved role for IP6 in lentivirus assembly. 著者: Robert A Dick / Chaoyi Xu / Dustin R Morado / Vladyslav Kravchuk / Clifton L Ricana / Terri D Lyddon / Arianna M Broad / J Ryan Feathers / Marc C Johnson / Volker M Vogt / Juan R Perilla / ...著者: Robert A Dick / Chaoyi Xu / Dustin R Morado / Vladyslav Kravchuk / Clifton L Ricana / Terri D Lyddon / Arianna M Broad / J Ryan Feathers / Marc C Johnson / Volker M Vogt / Juan R Perilla / John A G Briggs / Florian K M Schur / 要旨: Retrovirus assembly is driven by the multidomain structural protein Gag. Interactions between the capsid domains (CA) of Gag result in Gag multimerization, leading to an immature virus particle that ...Retrovirus assembly is driven by the multidomain structural protein Gag. Interactions between the capsid domains (CA) of Gag result in Gag multimerization, leading to an immature virus particle that is formed by a protein lattice based on dimeric, trimeric, and hexameric protein contacts. Among retroviruses the inter- and intra-hexamer contacts differ, especially in the N-terminal sub-domain of CA (CANTD). For HIV-1 the cellular molecule inositol hexakisphosphate (IP6) interacts with and stabilizes the immature hexamer, and is required for production of infectious virus particles. We have used in vitro assembly, cryo-electron tomography and subtomogram averaging, atomistic molecular dynamics simulations and mutational analyses to study the HIV-related lentivirus equine infectious anemia virus (EIAV). In particular, we sought to understand the structural conservation of the immature lentivirus lattice and the role of IP6 in EIAV assembly. Similar to HIV-1, IP6 strongly promoted in vitro assembly of EIAV Gag proteins into virus-like particles (VLPs), which took three morphologically highly distinct forms: narrow tubes, wide tubes, and spheres. Structural characterization of these VLPs to sub-4Å resolution unexpectedly showed that all three morphologies are based on an immature lattice with preserved key structural components, highlighting the structural versatility of CA to form immature assemblies. A direct comparison between EIAV and HIV revealed that both lentiviruses maintain similar immature interfaces, which are established by both conserved and non-conserved residues. In both EIAV and HIV-1, IP6 regulates immature assembly via conserved lysine residues within the CACTD and SP. Lastly, we demonstrate that IP6 stimulates in vitro assembly of immature particles of several other retroviruses in the lentivirus genus, suggesting a conserved role for IP6 in lentiviral assembly.
Tilt series were low-pass filtered according to their cumulative dose using exposure filters that were calculated using an exposure-dependent amplitude attenuation function and critical exposure constants (as published in Grant & Grigorieff, Elife, 2015). Tilt series were aligned and reconstructed in IMOD.
最終 再構成
アルゴリズム: BACK PROJECTION / ソフトウェア - 名称: IMOD / 使用した粒子像数: 41
CTF補正
ソフトウェア: (名称: CTFFIND (ver. 4), NOVACTF) / 詳細: CTF-correction was performed using NOVACTF