Protein–Protein Interactions Occurring During HIV Capsid Assembly in a Cell-free Protein Synthesizing System

Copeland, Kiel; Hansen, Wendy F.; Asundi, Vinod K.; Hong, Shi; Chamberlin, James W.; Dey, Debendranath; Broce, Sean; Himmel, Harold; Decloutte, Caleb; Ram, Salvin; Steffen, Imke; Pöhlmann, Stefan; Lingappa, Jaisri R.; Hurt, Clarence R.; Lingappa, Vishwanath R.

doi:10.1016/j.antiviral.2010.02.339

Cited by 1 publication

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“…Much of this prior work involved CFPSA systems programmed with mRNA encoding viral capsid proteins. A moderate throughput phenotypic drug screen involving CFPSA of influenza (FLUV) encoded proteins was developed (Petsch et al, 2010), analogous to what has also been done for rabies (Lingappa et al, 2013), HIV (Copeland et al, 2010; Reed et al, 2021), and other viruses (Broce et al, 2016). This screen is carried out in cellular extracts rather than in living cells, with formation of multimeric capsid protein complexes as a quantifiable, functional endpoint (Harrell, E.K.T.…”

Section: Resultsmentioning

confidence: 99%

“…Applied to drug discovery, this approach has identified small molecules termed assembly modulators, which have novel, host-targeted mechanisms of antiviral action. The host MPCs on which these compounds are active have been termed assembly machines 8 by virtue of their ability to catalyze capsid formation 10,8,11 Here we have utilized the same approach to study assembly of viruses implicated in severe respiratory disease. As a starting point, we began with FLUV, a member of family Orthomyxoviridae.…”

Section: Probing Viral Assembly With An Orthogonal Host-targeted Viramentioning

confidence: 99%

“…The host MPCs on which these compounds are active have been termed assembly machines by virtue of their ability to catalyze viral capsid assembly 10,8,11 .…”

Section: Probing Viral Assembly With An Orthogonal Host-targeted Viral Protein Biogenesis Screenmentioning

confidence: 99%

“…As a starting point, we began with FLUV, a member of the family Orthomyxoviridae. By analogy to previous studies on human immunodeficiency virus (HIV) 10,11 and rabies virus 8 , FLUV nucleoprotein (NP), the major capsid protein, was expressed together with other FLUV gene products including the matrix protein (M), in the CFPSA system. We observed a putative capsid assembly pathway (Figure 1A), roughly analogous to the pathways observed previously for other viral families, progression of whose assembly intermediates to the final structure is energy-dependent 8,9,12,[12][13][14][15] .…”

Section: Probing Viral Assembly With An Orthogonal Host-targeted Viral Protein Biogenesis Screenmentioning

confidence: 99%

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A Pan-Respiratory Antiviral Chemotype Targeting a Host Multi-Protein Complex

Selvarajah¹,

Lingappa²,

Michon³

et al. 2021

Preprint

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Antiviral compounds displaying remarkable features have been identified by an unconventional drug screen and advanced through animal validation. Efficacy is observed against the six viral families causing most human respiratory viral disease, irrespective of strain, including both influenza (FLUV) and SARS-CoV-2, with cell culture EC50 at or below 100 nM. Survival benefit is demonstrated in pigs against another member of family Coronaviridae, porcine epidemic diarrhea virus (PEDV), and shown equally effective in mild and severe disease. Respiratory syncytial virus (RSV) titer is reduced by drug treatment in cotton rats. A substantial barrier to viral resistance is demonstrated for FLUV. Drug resin affinity chromatography (DRAC) reveals a novel drug target: a multi-protein complex (MPC) formed transiently, in an energy-dependent fashion, and composed of host proteins implicated in both viral lifecycles and manipulation of innate immunity. The protein composition of this host MPC is modified upon viral infection, with increase or decrease of some proteins and appearance or complete loss of others. Valosin-containing protein, also known as Transitional endoplasmatic reticulum ATPase (VCP/p97), is present in the target MPC of uninfected cells and significantly increased in both FLUV and CoV infection. SQSTM1/p62, a key regulator of the autophagy pathway of innate immunity whose dysfunction is implicated in cytokine storm, is i) found in the target MPC from uninfected cells, ii) diminished in DRAC eluates by infection, and iii) restored by drug treatment of infected cells. 14-3-3 is one of likely several proteins that comprise the drug-binding site. Advanced compounds with improved pharmacokinetic (PK) properties and lung exposure are approaching criteria for a Target Product Profile. We propose these novel drug targets to comprise a previously unappreciated molecular basis for homeostasis that is modified by viruses to allow exploitation for viral propagation and is restored by treatment with the therapeutic compounds presented. This discovery has transformative implications for treatment of respiratory viral-related disease, applicable to everything from seasonal FLUV to COVID-19 and future novel respiratory viruses, due to the pan-family nature of drug activity and barrier to resistance development.

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Section: Resultsmentioning

confidence: 99%

Section: Probing Viral Assembly With An Orthogonal Host-targeted Viramentioning

confidence: 99%

“…The host MPCs on which these compounds are active have been termed assembly machines by virtue of their ability to catalyze viral capsid assembly 10,8,11 .…”

Section: Probing Viral Assembly With An Orthogonal Host-targeted Viral Protein Biogenesis Screenmentioning

confidence: 99%