Proteolytic cleavage of host proteins by the Group IV viral proteases of Venezuelan equine encephalitis virus and Zika virus

Morazzani, Elaine M.; Compton, Jaimee R.; Leary, Dagmar H.; Berry, Angela; Hu, Xin; Marugán, Juan; Glass, Pamela J.; Legler, Patricia M.

doi:10.1016/j.antiviral.2019.02.001

Cited by 24 publications

(53 citation statements)

References 114 publications

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“…However, at longer periods, the cleavage products were not detected. In virus-infected cells, the kinetics were different and cleavage products could be detected between 6-48 h 6 . Others have reported the appearance of host protein cleavage products as early as 3-6 h post infection 9,11 .…”

Section: Introductionmentioning

confidence: 98%

“…These sequences within the coding regions of the viral genome can be propagated generationally. The short stretches of homologous host-pathogen protein sequences (SSHHPS) are found in the virus and host 5,6 . SSHHPS are the conserved cleavage site motif sequences recognized by viral proteases that have homology to specific host proteins.…”

Section: Introductionmentioning

confidence: 99%

“…In our previous publication 6 , we compiled a list of all of the host proteins that were targeted by viral proteases and found that the list of targets was non-random ( Table 1). Two trends were apparent.…”

Section: Introductionmentioning

confidence: 99%

“…Sequence alignments of the host proteins targeted by viral proteases have revealed species-specific differences within these short cleavage site sequences. For example, the Venezuelan equine encephalitis virus (VEEV) nsP2 protease was found to cut human TRIM14, a tripartite motif (TRIM) protein 6 . Some TRIM proteins are viral restriction factors (e.g., TRIM5α 21 ), most are thought to be ubiquitin E3 ligases.…”

Section: Introductionmentioning

confidence: 99%

“…Cell-based assays can be valuable for establishing a role for the identified host protein(s) in infection. Methods that aim to halt viral protease cleavage of host proteins such as the addition of a protease inhibitor 6 or a mutation in the host target 16 can be used to examine their effects on viral replication. Overexpression of the targeted protein also may affect viral replication 28 .…”

Section: Introductionmentioning

confidence: 99%

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Analysis of Group IV Viral SSHHPS Using In Vitro and In Silico Methods

Compton

Legler

2019

JoVE

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Alphaviral enzymes are synthesized in a single polypeptide. The nonstructural polyprotein (nsP) is processed by its nsP2 cysteine protease to produce active enzymes essential for viral replication. Viral proteases are highly specific and recognize conserved cleavage site motif sequences (~6-8 amino acids). In several Group IV viruses, the nsP protease(s) cleavage site motif sequences can be found in specific host proteins involved in generating the innate immune responses and, in some cases, the targeted proteins appear to be linked to the virus-induced phenotype. These viruses utilize short stretches of homologous host-pathogen protein sequences (SSHHPS) for targeted destruction of host proteins. To identify SSHHPS the viral protease cleavage site motif sequences can be inputted into BLAST and the host genome(s) can be searched. Cleavage initially can be tested using the purified nsP viral protease and fluorescence resonance energy transfer (FRET) substrates made in E. coli. The FRET substrates contain cyan and yellow fluorescent protein and the cleavage site sequence (CFP-sequence-YFP). This protease assay can be used continuously in a plate reader or discontinuously in SDS-PAGE gels. Models of the bound peptide substrates can be generated in silico to guide substrate selection and mutagenesis studies. CFP/YFP substrates have also been utilized to identify protease inhibitors. These in vitro and in silico methods can be used in combination with cell-based assays to determine if the targeted host protein affects viral replication.

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Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%