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Human heart cell proteins interacting with a C-terminally truncated 2A protein of coxsackie B3 virus: identification by the yeast two-hybrid system

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Abstract

Protein 2A is a non-structural protein of coxsackievirus B3 (CVB3), an important human pathogen that can cause a variety of human diseases. Protein 2A not only participates in viral life cycle, but also regulates host cell functions; however, the underlying mechanisms remain poorly understood. In order to better understand the molecular mechanisms of CVB3 2A’s function, the yeast two-hybrid (Y2H) system was adopted to screen for CVB3 2A interactive proteins in the human heart cDNA library. Full-length 2A shows strong transcriptional activity in yeast cells, which interferes with the application of Y2H system; therefore, a series of 2A deletion mutants were constructed. Analysis of transcriptional self-activation revealed that 2A lost its transcriptional activity after truncation of 60 amino acids (aa) at the N-terminus or deletion of 17 aa at the C-terminus. Choosing the 2A mutant with 17 aa deletion at the C-terminus as the bait protein, four interactive cellular proteins were identified, including TIMP4, MYL2, COX7C, and ENO1. These proteins are mostly related to protein degradation and metabolism. Although the interactions detected by the Y2H system should be considered as preliminary results, the finding of proteins translated from a human heart cDNA library that interacts with the CVB3 2A will stimulate experiments testing the reactivity of a translational mixture derived from that library with full-length 2A protein, followed by co-immunoprecipitation studies.

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Acknowledgments

This work was supported by the Natural Science Foundation of Jiangxi Province in China (20142BAB205070) and National Natural Science Foundation of China (30660010 and 81160196).

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Authors and Affiliations

  1. School of Pharmacy, Nanchang University, Nanchang, 330006, Jiangxi, China

    Tiansheng Zhao

  2. Department of Medical Microbiology, School of Medicine, Nanchang University, Nanchang, 330006, Jiangxi, China

    Xiaotian Huang & Yanhua Xia

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  1. Tiansheng Zhao
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  2. Xiaotian Huang
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  3. Yanhua Xia
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Correspondence to Yanhua Xia.

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Edited by Zhen F. Fu.

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Zhao, T., Huang, X. & Xia, Y. Human heart cell proteins interacting with a C-terminally truncated 2A protein of coxsackie B3 virus: identification by the yeast two-hybrid system. Virus Genes 52, 172–178 (2016). https://doi.org/10.1007/s11262-015-1270-1

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