New World alphavirus protein interactomes from a therapeutic perspective

Carey, Brian D.; Bakovic, Allison; Callahan, Victoria; Narayanan, Aarthi

doi:10.1016/j.antiviral.2019.01.015

Cited by 22 publications

(21 citation statements)

References 139 publications

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“…We next sought to determine the step or steps in the viral replication cycle at which the mutants 204/5, 142, and 134/5 in VEEV nsP3 negatively affected viral replication. VEEV replication begins after viral entry and uncoating of the genome, following which the positive-sense genomic RNA is translated into the non-structural polyprotein and together, with various host proteins, the replication complex binds to the 3′ conserved sequence element (CSE) on the genomic RNA to initiate synthesis of the negative RNA strand [9,51]. Subsequent cleavages of the non-structural polyprotein into its constituents (nsP1-4) initiate positive-sense RNA synthesis of the full-length genome (49S) and subgenome (26S) via promoters on the negative-strand [51].…”

Section: Residues 204/5 142 and 134/5 Are Phosphorylated By Ikkβ Acmentioning

confidence: 99%

“…The nsP3 of both Old World and New World alphaviruses has been studied due to its capacity for hyperphosphorylation, variability among alphaviruses, and its interaction with a variety of host proteins [9,12]. However, cellular kinases responsible for phosphorylating various sites in nsP3 have yet to be determined [12].…”

Section: The Role Of Ikkβ During the Alphavirus Replication Cyclementioning

confidence: 99%

“…The VEEV genome consists of a positive-sense, single stranded RNA molecule~11.4 kb in length that encodes two polyproteins [1,5]. The non-structural polyprotein consists of the four non-structural proteins nsP1-4, that drive viral RNA replication in concert with a range of host proteins [9]. The structural polyprotein is cleaved into the viral capsid and glycoproteins, which are responsible for packaging genomes into progeny virions [1,10].…”

Section: Introductionmentioning

confidence: 99%

“…The structural polyprotein is cleaved into the viral capsid and glycoproteins, which are responsible for packaging genomes into progeny virions [1,10]. While multiple activities of nsP1, 2, and 4 have been described, the activities of nsP3 have only recently begun to be elucidated [9]. Three distinct domains in nsP3 have been categorized; an N-terminal macrodomain conserved among many alphaviruses, which exhibits homology with other RNA viruses, an alphavirus unique domain (AUD), and a C-terminal hypervariable domain (HVD) [11,12].…”

Section: Introductionmentioning

confidence: 99%

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Venezuelan Equine Encephalitis Virus nsP3 Phosphorylation Can Be Mediated by IKKβ Kinase Activity and Abrogation of Phosphorylation Inhibits Negative-Strand Synthesis

Bakovic

Bhalla

Kortchak

et al. 2020

Viruses

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Venezuelan equine encephalitis virus (VEEV), a mosquito transmitted alphavirus of the Togaviridae family, can cause a highly inflammatory and encephalitic disease upon infection. Although a category B select agent, no FDA-approved vaccines or therapeutics against VEEV currently exist. We previously demonstrated NF-κB activation and macromolecular reorganization of the IKK complex upon VEEV infection in vitro, with IKKβ inhibition reducing viral replication. Mass spectrometry and confocal microscopy revealed an interaction between IKKβ and VEEV non-structural protein 3 (nsP3). Here, using western blotting, a cell-free kinase activity assay, and mass spectrometry, we demonstrate that IKKβ kinase activity can directly phosphorylate VEEV nsP3 at sites 204/5, 142, and 134/5. Alanine substitution mutations at sites 204/5, 142, or 134/5 reduced VEEV replication by >30-100,000-fold corresponding to a severe decrease in negative-strand synthesis. Serial passaging rescued viral replication and negative-strand synthesis, and sequencing of revertant viruses revealed reversion to the wild-type TC-83 phosphorylation capable amino acid sequences at nsP3 sites 204/5, 142, and 135. Generation of phosphomimetic mutants using aspartic acid substitutions at site 204/5 resulted in rescue of both viral replication and negative-strand RNA production, whereas phosphomimetic mutant 134/5 rescued viral replication but failed to restore negative-strand RNA levels, and phosphomimetic mutant 142 did not rescue VEEV replication. Together, these data demonstrate that IKKβ can phosphorylate VEEV nsP3 at sites 204/5, 142, and 134/5, and suggest that phosphorylation is essential for negative-strand RNA synthesis at site 204/5, but may be important for infectious particle production at site 134/5.

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Section: Residues 204/5 142 and 134/5 Are Phosphorylated By Ikkβ Acmentioning

confidence: 99%

Section: The Role Of Ikkβ During the Alphavirus Replication Cyclementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Venezuelan Equine Encephalitis Virus nsP3 Phosphorylation Can Be Mediated by IKKβ Kinase Activity and Abrogation of Phosphorylation Inhibits Negative-Strand Synthesis

Bakovic

Bhalla

Kortchak

et al. 2020

Viruses

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“…Humans can be infected by spillover of virus, resulting in disease that is similar to but milder than RRV infection, having relatively benign symptoms such as fever, rash, arthralgia, and myalgia (2–5). There is no specific antiviral treatment, and no commercial BFV vaccine is available (2, 6, 7).…”

Section: Announcementmentioning

confidence: 99%

Genome Sequences of Barmah Forest Virus Strains Isolated from Mosquitoes Trapped in Australian Defence Force Training Areas Reveal Multiple Nucleotide Insertions in the 3′ Untranslated Region

Liu¹,

Kizu²,

Hoare³

et al. 2019

Microbiol Resour Announc

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Self-amplifying mRNA-Based Vaccine Technology and Its Mode of Action

Maruggi

Ulmer

Rappuoli³

et al. 2021

Current Topics in Microbiology and Immunology

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Self-amplifying mRNAs derived from the genomes of positive-strand RNA viruses have recently come into focus as a promising technology platform for vaccine development. Non-virally delivered self-amplifying mRNA vaccines have the potential to be highly versatile, potent, streamlined, scalable, and inexpensive. By amplifying their genome and the antigen encoding mRNA in the host cell, the self-amplifying mRNA mimics a viral infection, resulting in sustained levels of the target protein combined with self-adjuvanting innate immune responses, ultimately leading to potent and long-lasting antigen-specific humoral and cellular immune responses. Moreover, in principle, any eukaryotic sequence could be encoded by self-amplifying mRNA without the need to change the manufacturing process, thereby enabling a much faster and flexible research and development timeline than the current vaccines and hence a quicker response to emerging infectious diseases. This chapter highlights the rapid progress made in using non-virally delivered self-amplifying mRNA-based vaccines against infectious diseases in animal models. We provide an overview of the unique attributes of this vaccine approach, summarize the growing body of work defining its mechanism of action, discuss the current challenges and latest advances, and highlight perspectives about the future of this promising technology.

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New World alphavirus protein interactomes from a therapeutic perspective

Cited by 22 publications

References 139 publications

Venezuelan Equine Encephalitis Virus nsP3 Phosphorylation Can Be Mediated by IKKβ Kinase Activity and Abrogation of Phosphorylation Inhibits Negative-Strand Synthesis

Venezuelan Equine Encephalitis Virus nsP3 Phosphorylation Can Be Mediated by IKKβ Kinase Activity and Abrogation of Phosphorylation Inhibits Negative-Strand Synthesis

Genome Sequences of Barmah Forest Virus Strains Isolated from Mosquitoes Trapped in Australian Defence Force Training Areas Reveal Multiple Nucleotide Insertions in the 3′ Untranslated Region

Self-amplifying mRNA-Based Vaccine Technology and Its Mode of Action

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