Viral–Host Protein Interaction Studies Using Yeast Two-Hybrid Screening Method

Dudha, Namrata; Gupta, Sanjay

doi:10.1007/978-1-4939-3618-2_15

Cited by 1 publication

(1 citation statement)

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“…Thus, it is imperative to identify the novel cellular proteins interacting with 2C for further insights into the virus replication and pathogenesis. Yeast two-hybrid (Y2H) is one of the most versatile and high throughput tools available to study virus-host proteinprotein interaction and has been used in several studies earlier [13,14]. In this study, we have also employed the Y2H system to identify the 2C-interacting host proteins.…”

Section: Introductionmentioning

confidence: 99%

Identification of novel interactions between host and non-structural protein 2C of foot-and-mouth disease virus

Mahajan

Sharma

Bora

et al. 2021

Journal of General Virology

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The 2C protein of foot-and-mouth disease virus (FMDV) is reported to play a critical role in the virus replication complex and modulating the host’s immune response. However, the underlying molecular intricacies of subversion of cellular machinery remains poorly understood, thus emphasizing the need to study 2C-host interactions. In this study, we identified the host proteins interacting with the 2C using yeast-two hybrid (Y2H) approach, which is one of the most recognized, high-throughput tools to study protein-protein interactions. The FMDV-2C bait was characterized for auto-activation, toxicity, and expression and was found to be suitable for mating with cDNA library. On preliminary screening a total of 32 interacting host proteins were identified which were reduced to 22 on subsequent confirmation with alternative yeast based assays. Amongst these, NMI/2C interaction has been reported earlier by Wang et al. (2012) and remaining 21 are novel interactions. The Reactome analysis has revealed the role of the identified host proteins in cellular pathways exploited by 2C during FMDV replication. We also confirmed interaction of MARCH7, an E3 ubiquitin ligase with 2C using mammalian two-hybrid system and co-immunoprecipitation. This study leads to the identification of novel 2C interacting host proteins which enhance our understanding of 2C-host interface and may provide checkpoints for development of potential therapeutics against FMDV.

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Section: Introductionmentioning

confidence: 99%