
抗体設計において、ウイルスの進化を予測し、その進化を抑制する抗体を設計することが重要である。


coremsg

ウイルス進化を予測し抗体を設計する

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ZIKV ウイルスは IGF2BP2 リボ核タンパク質複合体を改変し乗っ取ることで、ウイルス複製オルガネラの形成を促進する


Zika virus (ZIKV) infection causes significant human disease that, with no approved treatment or vaccine, constitutes a major public health concern. Its life cycle entirely relies on the cytoplasmic fate of the viral RNA genome (vRNA) through a fine-tuned equilibrium between vRNA translation, replication and packaging into new virions, all within virus-induced replication organelles (vRO). In this study, with an RNAi mini-screening and subsequent functional characterization, we have identified insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) as a new host dependency factor that regulates vRNA synthesis. In infected cells, IGF2BP2 associates with viral NS5 polymerase and redistributes to the perinuclear viral replication compartment. Combined fluorescence in situ hybridization-based confocal imaging, in vitro binding assays, and immunoprecipitation coupled to RT-qPCR, showed that IGF2BP2 directly interacts with ZIKV vRNA 3’-nontranslated region. Using ZIKV sub-genomic replicons and a replication-independent vRO induction system, we demonstrated that IGF2BP2 knockdown impairs de novo viral organelle biogenesis and, consistently, vRNA synthesis. Finally, the analysis of immunopurified IGF2BP2 complex using quantitative mass spectrometry and RT-qPCR, revealed that ZIKV infection alters the protein and RNA interactomes of IGF2BP2. Altogether, our data support that ZIKV hijacks and remodels the IGF2BP2 ribonucleoprotein complex to regulate vRO biogenesis and vRNA neosynthesis.

### Competing Interest Statement

The authors have declared no competing interest.

Zika virus remodels and hijacks IGF2BP2 ribonucleoprotein complex to promote viral replication organelle biogenesis

zikv-ウイルスは-igf2bp2-リボ核タンパク質複合体を改変し乗っ取ることで-ウイルス複製オルガネラの形成を促進する


HIVエリートコントローラーにおいて、特定の逆転写可能な要素の発現上昇が、抗ウイルス遺伝子の発現を高めることで、HIVに対する抵抗性を高めている可能性がある。


Less than 0.5% of people living with HIV-1 are elite controllers (ECs) - individuals who have a replication-competent viral reservoir in their CD4+ T cells but maintain undetectable plasma viremia without the help of antiretroviral therapy. While the EC CD4+ T cell transcriptome has been investigated for gene expression signatures associated with disease progression (or, in this case, a lack thereof), the expression and regulatory activity of transposable elements (TEs) in ECs has not been explored. Yet previous studies have established that TEs can directly impact the immune response to pathogens, including HIV-1. Thus, we hypothesize that the regulatory activities of TEs could contribute to the natural resistance of ECs against HIV-1. We perform a TE-centric analysis of previously published multi-omics data derived from EC individuals and other populations. We find that the CD4+ T cell transcriptome and retrotranscriptome of ECs are distinct from healthy controls, treated patients, and viremic progressors. However, there is a substantial level of transcriptomic heterogeneity among ECs. We categorize individuals with distinct chromatin accessibility and expression profiles into four clusters within the EC group, each possessing unique repertoires of TEs and antiviral factors. Notably, several TE families with known immuno-regulatory activity are differentially expressed among ECs. Their transcript levels in ECs positively correlate with their chromatin accessibility and negatively correlate with the expression of their KRAB zinc-finger (KZNF) repressors. This coordinated variation is seen at the level of individual TE loci likely acting or, in some cases, known to act as cis -regulatory elements for nearby genes involved in the immune response and HIV-1 restriction. Based on these results, we propose that the EC phenotype is driven in part by the reduced availability of specific KZNF proteins to repress TE-derived cis -regulatory elements for antiviral genes, thereby heightening their basal level of resistance to HIV-1 infection. Our study reveals considerable heterogeneity in the CD4+ T cell transcriptome of ECs, including variable expression of TEs and their KZNF controllers, that must be taken into consideration to decipher the mechanisms enabling HIV-1 control.

### Competing Interest Statement

The authors have declared no competing interest.

Transposable elements may enhance antiviral resistance in HIV-1 elite controllers

hivエリートコントローラーにおける逆転写可能な要素による抗ウイルス抵抗性の強化


HIVエリートコントローラーにおける逆転写可能な要素による抗ウイルス抵抗性の強化



SARS-CoV-2感染によりホスト細胞でYAPの転写活性が抑制される。SARS-CoV-2のNSP13タンパク質がTEAD4と結合し、YAP/TEAD4複合体を不活性化することで、YAPの転写活性を抑制する。


The Hippo pathway is critical to organ development, homeostasis, and regeneration, facilitated by YAP/TEAD-mediated gene expression. Although emerging studies report Hippo-YAP dysfunction after viral infection, it is largely unknown in the context of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here, we analyzed RNA sequencing data from SARS-CoV-2 infected human lung samples and induced pluripotent stem cell–derived cardiomyocytes (iPSC-CMs), and observed a decrease in YAP target gene expression. In screening SARS-CoV-2 nonstructural proteins, we found that nonstructural protein 13 (NSP13), a conserved coronavirus helicase, inhibited YAP transcriptional activity independent of the upstream Hippo kinases LATS1/2. Consistently, introducing NSP13 into cardiomyocytes suppressed active YAP (YAP5SA) in vivo . Subsequent investigations on NSP13 mutants indicated that NSP13 helicase activity is crucial for suppressing YAP transactivation. Mechanistically, TEAD4 serves as a platform for recruiting NSP13 and YAP. NSP13 inactivates the YAP/TEAD4 transcription complex through its interacting proteins, such as transcription termination factor 2 (TTF2). These discoveries reveal a novel YAP/TEAD regulatory mechanism orchestrated by TEAD4, which provides molecular insights of Hippo-YAP regulation after SARS-CoV-2 infection.

### Competing Interest Statement

J.F.M is a founder and owns shares in Yap therapeutics. J.F.M. is a co-inventor on the following patents associated with this study: patent no. US20200206327A1 entitled Hippo pathway deficiency reverses systolic heart failure post-infarction, patent no.15/642200.PCT/US2014/069349 101191411 entitled Hippo and dystrophin complex signaling in cardiomyocyte renewal, and patent no. 15/102593.PCT/US2014/069349 9732345 entitled Hippo and dystrophin complex signaling in cardiomyocyte renewal.

SARS-CoV-2 NSP13 interacts with TEAD to suppress Hippo-YAP signaling

sars-cov-2のnsp13タンパク質がhippoシグナル経路の下流エフェクターyapを抑制する


SARS-CoV-2のNSP13タンパク質がHippoシグナル経路の下流エフェクターYAPを抑制する



Malectinはコロナウイルス感染を促進し、ウイルス蛋白質の生合成を支援する。


Coronaviruses (CoV) rewire host protein homeostasis (proteostasis) networks through interactions between viral nonstructural proteins (nsps) and host factors to promote infection. With the emergence of SARS-CoV-2, it is imperative to characterize host interactors shared across nsp homologs. Using quantitative proteomics and functional genetic screening, we identify conserved proteostasis interactors of nsp2 and nsp4 that serve pro-viral roles during infection of murine hepatitis virus – a model betacoronavirus. We uncover a glycoprotein quality control factor, Malectin (MLEC), which significantly reduces infectious titers when knocked down. During infection, nsp2 interacts with MLEC-associated proteins and the MLEC-interactome is drastically altered, stabilizing association with the Oligosaccheryltransferase (OST) complex, a crucial component of viral glycoprotein production. MLEC promotes viral protein levels and genome replication through its quality control activity. Lastly, we show MLEC promotes SARS-CoV-2 replication. Our results reveal a role for MLEC in mediating CoV infection and identify a potential target for pan-CoV antivirals.

### Competing Interest Statement

The authors have declared no competing interest.

The mass spectrometry proteomics data are deposited to the ProteomeXchange Consortium via the PRIDE partner repository under the accession code PXD049130. All other necessary data are contained within the manuscript or can be shared by the Lead Contact upon request.

The glycoprotein quality control factor Malectin promotes coronavirus replication and viral protein biogenesis

コロナウイルス複製とウイルス蛋白質生合成を促進するグリコ蛋白質品質管理因子malectin


コロナウイルス複製とウイルス蛋白質生合成を促進するグリコ蛋白質品質管理因子Malectin



KSHV感染細胞では、ウイルス複製に必須の BiP/GRP78が UPR非依存的に発現が上昇し、ウイルス増殖を促進する。BiP/GRP78の阻害は、KSHV関連悪性腫瘍の治療の可能性を示唆する。


The Endoplasmic Reticulum (ER)-resident HSP70 chaperone BiP (HSPA5) plays a crucial role in maintaining and restoring protein folding homeostasis in the ER. BiP’s function is often dysregulated in cancer and virus-infected cells, conferring pro-oncogenic and pro-viral advantages. We explored BiP’s functions during infection by the Kaposi’s sarcoma-associated herpesvirus (KSHV), an oncogenic gamma-herpesvirus associated with cancers of immunocompromised patients. Our findings reveal that BiP protein levels are upregulated in infected epithelial cells during the lytic phase of KSHV infection. This upregulation occurs independently of the unfolded protein response (UPR), a major signaling pathway that regulates BiP availability. Genetic and pharmacological inhibition of BiP halts KSHV viral replication and reduces the proliferation and survival of KSHV-infected cells. Notably, inhibition of BiP limits the spread of other alpha- and beta-herpesviruses and poxviruses with minimal toxicity for normal cells. Our work suggests that BiP is a potential target for developing broad-spectrum antiviral therapies against double-stranded DNA viruses and a promising candidate for therapeutic intervention in KSHV-related malignancies.

### Competing Interest Statement

The authors have declared no competing interest.

BiP/GRP78 is a pro-viral factor for diverse dsDNA viruses that promotes the survival and proliferation of cells upon KSHV infection

kshv感染細胞における生存と増殖を促進する-pro-viral-因子としての-bip-grp78


KSHV感染細胞における生存と増殖を促進する pro-viral 因子としての BiP/GRP78



動物サーベコウイルスの一部は、ACE2非依存的かつトリプシン依存的な細胞侵入経路を利用し、これにより抗体介在性中和に対する感受性が低下する。


The COVID-19 pandemic, caused by SARS-CoV-2, demonstrated that zoonotic transmission of animal sarbecoviruses threatens human health but the determinants of transmission are incompletely understood. Here, we show that most spike (S) proteins of horseshoe bat and Malayan pangolin sarbecoviruses employ ACE2 for entry, with human and raccoon dog ACE2 exhibiting broad receptor activity. The insertion of a multibasic cleavage site into the S proteins increased entry into human lung cells driven by most S proteins tested, suggesting that acquisition of a multibasic cleavage site might increase infectivity of diverse animal sarbecoviruses for the human respiratory tract. In contrast, two bat sarbecovirus S proteins drove cell entry in an ACE2-independent, trypsin-dependent fashion and several ACE2-dependent S proteins could switch to the ACE2-independent entry pathway when exposed to trypsin. Several TMPRSS2-related cellular proteases but not the insertion of a multibasic cleavage site into the S protein allowed for ACE2-independent entry in the absence of trypsin and may support viral spread in the respiratory tract. Finally, the pan-sarbecovirus antibody S2H97 enhanced cell entry driven by two S proteins and this effect was reversed by trypsin. Similarly, plasma from quadruple vaccinated individuals neutralized entry driven by all S proteins studied, and use of the ACE2-independent, trypsin-dependent pathway reduced neutralization sensitivity. In sum, our study reports a pathway for entry into human cells that is ACE2-independent, supported by TMPRSS2-related proteases and associated with antibody evasion.

### Competing Interest Statement

S.P. and M.H. conducted contract research (testing of vaccinee plasma for neutralizing activity against SARS-CoV-2) for Valneva unrelated to this work. G.M.N.B. served as advisor for Moderna and S.P. served as advisor for BioNTech, unrelated to this work. The authors declare no competing interests. M.S.W. received funding from Sartorius AG (Goettingen, Germany) from GRIFOLS SA (Barcelona, Spain), Sphingotec (Henningsdorf, Germany), Inflammatix (Sunnyvale, CA, USA) and the German Research Foundation (Bonn, Germany). He is in the advisory board of Amomed (Wien, Austria) and Gilead Science Inc. (Foster City, CA, USA).

ACE2-independent sarbecovirus cell entry is supported by TMPRSS2-related enzymes and reduces sensitivity to antibody-mediated neutralization

動物サーベコウイルスのace2非依存的な細胞侵入は-tmprss2関連酵素によってサポートされ-抗体介在性中和に対する感受性を低下させる


動物サーベコウイルスのACE2非依存的な細胞侵入は、TMPRSS2関連酵素によってサポートされ、抗体介在性中和に対する感受性を低下させる



グレーオオカミにおいて高頻度で検出されるキツネサーコウイルスは高い遺伝的多様性を示し、野生動物における重要な保菌宿主であることが示唆された。


Canine circoviruses (CanineCV) have a worldwide distribution in dogs and are occasionally detected in wild carnivorans, indicating their ability for cross-species transmission. However, fox circovirus, a lineage of CanineCV, has been identified exclusively in wild canids. We analyzed spleen samples from 159 grey wolves from the Northwest Territories, Canada, to investigate the molecular epidemiology of CanineCV and formulate hypotheses about virus ecology and evolution. Overall, 72 out of 159 (45.3%) animals tested positive. Virus prevalence was similar between males and females, adults and juveniles, and across the investigated years and locations. CanineCV infection was not associated with a poor body condition. While the percentage of co-infections with canine parvoviruses, investigated in a previous study, was high (63/72, 87.5%), the rate of parvovirus infection in CanineCV-negative animals was significantly lower (42/87, 48.3%, χ2 = 27.03, p < 0.001), and CanineCV infection was associated with a 7.5- and 2.4-fold increase in the risk of acquiring canine parvovirus 2 or canine bufavirus infections, respectively (odds ratios: 3.5-16.9 and 1.3-5.8). Although common risk factors cannot be ruled out, this suggests that CanineCV may facilitate parvoviral super-infections. Sequencing revealed high CanineCV genetic diversity, further exacerbated by recombination. Of the 69 sequenced strains, 87.5% were fox circoviruses, five were related to a fox circovirus-like recombinant strain, and one belonged to a distant lineage. In the phylogenetic analysis, the virus sequences were distributed according to sampling locations, with some viruses being geographically restricted. Different clades of viruses were identified in the same areas and over multiple years (2007-2019), indicating the co-existence of multiple endemic lineages in the investigated area. Phylogenetic analysis of all available complete fox circovirus genomes (32 from foxes and 15 from wolves from North America and Europe) demonstrated four lineages, each including sequences from this study. Within each lineage, strains segregated geographically and not by host. This implies that, although multiple lineages co-exist, viruses do not frequently move between locations. Finally, viruses from Europe and North America were mixed, indicating that the origin of the four lineages might predate the segregation of European and American wolf and fox populations. Given the high prevalence and diversity of fox circoviruses in wolves, these animals should be considered reservoir hosts for these viruses. Although we cannot exclude a lower susceptibility of dogs, the lack of fox circovirus in dogs could be due to environmental circumstances that prevented its spread to dogs. Given the high diversity and wild host specificity, we presume a long-lasting association between fox circovirus and canine hosts and hypothesize a higher likelihood of transmission from dogs to wild animals than vice versa. Further studies should investigate other sympatric wild species and additional locations to explore the possible existence of additional maintenance hosts and the reasons behind the marked difference in cross-species transmission dynamics among CanineCV lineages.

### Competing Interest Statement

The authors have declared no competing interest.

Diverse fox circovirus (Circovirus canine) variants circulate at high prevalence in grey wolves (Canis lupus) from the Northwest Territories, Canada

北米のグレーオオカミ-canis-lupus-から高頻度で検出される多様なキツネサーコウイルス-circovirus-canine-変異株


北米のグレーオオカミ(Canis lupus)から高頻度で検出される多様なキツネサーコウイルス(Circovirus canine)変異株



HBVコアタンパク質の二つの非従来的な結合部位を標的とした合成ダイマー化合物が、コアタンパク質の凝集を引き起こすことが明らかになった。


The hepatitis B virus (HBV) infection is a major global health problem, with chronic infection leading to liver complications and high death toll. Current treatments, such as nucleos(t)ide analogs and interferon-α, effectively suppress viral replication but rarely cure the infection. To address this, new antivirals targeting different components of the HBV molecular machinery are being developed. Here we investigated the hepatitis B core protein (HBc) that forms the viral capsids and plays a vital role in the HBV life cycle. We explored two distinct binding pockets on the HBV capsid: the central hydrophobic pocket of HBc-dimers and the pocket at the tips of capsid spikes. We synthesized a geranyl dimer that binds to the central pocket with micromolar affinity, and dimeric peptides that bind the spike-tip pocket with nanomolar affinity. Cryo-electron microscopy further confirmed the binding of peptide dimers to the capsid spike tips and their capsid-aggregating properties. Finally, we show that the peptide dimers induce HBc aggregation in vitro and in living cells. Our findings highlight two tractable sites within the HBV capsid and provide an alternative strategy to affect HBV capsids.

### Competing Interest Statement

The authors have declared no competing interest.

Induction of Hepatitis B Core Protein Aggregation Targeting an Unconventional Binding Site

hbvコアタンパク質の凝集を標的とした非従来的な結合部位の誘導


HBVコアタンパク質の凝集を標的とした非従来的な結合部位の誘導



HIVウイルス-1は宿主細胞のR-ループを誘導し、それらのR-ループ豊富な領域に選好的に統合する。


Although HIV-1 integration sites are considered to favor active transcription units in the human genome, high-resolution analysis of individual HIV-1 integration sites have shown that the virus can integrate in a variety of host genomic locations, including non-genic regions. The invisible infection by HIV-1 integrating into non-genic regions challenging the traditional understanding of HIV-1 integration site selection are rather more problematic as they are selected to preserve in the host genome during prolonged antiretroviral therapies. Here, we showed that HIV-1 targets R-loops, a genomic structure made up of DNA–RNA hybrids, for integration. HIV-1 initiates the formation of R-loops in both genic and non-genic regions of the host genome and preferentially integrates into R-loop-rich regions. Using a cell model that can independently control transcriptional activity and R-loop formation, we demonstrated that the formation of R-loops directs HIV-1 integration targeting sites. We also found that HIV-1 integrase proteins physically bind to the host genomic R-loops. These findings provide fundamental insights into the mechanisms of retroviral integration and the new strategies of antiretroviral therapy against HIV-1 latent infection.

### Competing Interest Statement

The authors have declared no competing interest.

Human immunodeficiency virus-1 induces and targets host genomic R-loops for viral genome integration

hivウイルス-1は宿主ゲノムのr-ループを誘導し-ウイルスゲノム統合のためのターゲットとする


HIVウイルス-1は宿主ゲノムのR-ループを誘導し、ウイルスゲノム統合のためのターゲットとする
