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EV71 5'UTR interacts with 3D protein affecting replication through the AKT-mTOR pathway

Xiaoying Xu¹, Shao Ma², Ziwei Liu³

¹School of Public Health, Cheeloo College of Medicine, Shandong University, No. 44 Wenhua West Road, Lixia District, Jinan, 250012, China.

Virology Journal

|May 22, 2024

Abstract

Background:

EV71 is one of the important pathogens of Hand-foot-and-mouth disease (HFMD), which causes serious neurological symptoms. Several studies have speculated that there will be interaction between 5'UTR and 3D protein. However, whether 5'UTR interacts with the 3D protein in regulating virus replication has not been clarified.

Methods:

Four 5'UTR mutation sites (nt88C/T, nt90-102-3C, nt157G/A and nt574T/A) and two 3D protein mutation sites (S37N and R142K) were mutated or co-mutated using virulent strains as templates. The replication of these mutant viruses and their effect on autophagy were determined.

Results:

5'UTR single-point mutant strains, except for EGFP-EV71(nt90-102-3C), triggered replication attenuation. The replication ability of them was weaker than that of the parent strain the virulent strain SDLY107 which is the fatal strain that can cause severe neurological complications. While the replication level of the co-mutant strains showed different characteristics. 5 co-mutant strains with interaction were screened: EGFP-EV71(S37N-nt88C/T), EGFP-EV71(S37N-nt574T/A), EGFP-EV71(R142K-nt574T/A), EGFP-EV71(R142K-nt88C/T), and EGFP-EV71(R142K-nt157G/A). The results showed that the high replicative strains significantly promoted the accumulation of autophagosomes in host cells and hindered the degradation of autolysosomes. The low replicative strains had a low ability to regulate the autophagy of host cells. In addition, the high replicative strains also significantly inhibited the phosphorylation of AKT and mTOR.

Conclusions:

EV71 5'UTR interacted with the 3D protein during virus replication. The co-mutation of S37N and nt88C/T, S37N and nt574T/ A, R142K and nt574T/A induced incomplete autophagy of host cells and promoted virus replication by inhibiting the autophagy pathway AKT-mTOR. The co-mutation of R142K and nt88C/T, and R142K and nt157G/A significantly reduced the inhibitory effect of EV71 on the AKT-mTOR pathway and reduced the replication ability of the virus.

Insights

This study reveals that Enterovirus 71 (EV71) 5'UTR interacts with the 3D protein to regulate virus replication. Specific mutations in EV71

Area of Science:

Virology
Molecular Biology
Cellular Biology

Background:

Enterovirus 71 (EV71) is a significant cause of Hand-foot-and-mouth disease (HFMD), often leading to severe neurological complications.
Previous research suggested a potential interaction between the EV71 5' untranslated region (5'UTR) and the 3D protein, but its role in viral replication remained unclear.

Purpose of the Study:

To investigate the interaction between the EV71 5'UTR and the 3D protein.
To determine the impact of this interaction on viral replication and host cell autophagy.

Main Methods:

Mutagenesis of four 5'UTR sites and two 3D protein sites in virulent EV71 strains.
Analysis of mutant virus replication and their effects on cellular autophagy.
Assessment of the autophagy pathway, including autophagosome accumulation, autolysosome degradation, and AKT/mTOR signaling.

Main Results:

Single 5'UTR mutations, except for one, attenuated viral replication compared to the virulent parent strain.
Co-mutant strains exhibited varied replication characteristics, with five showing evidence of interaction.
High-replicative strains promoted autophagosome accumulation, hindered autolysosome degradation, and inhibited AKT/mTOR phosphorylation.
Low-replicative strains showed minimal regulation of host cell autophagy.

Conclusions:

The EV71 5'UTR interacts with the 3D protein, influencing viral replication.
Specific co-mutations (S37N-nt88C/T, S37N-nt574T/A, R142K-nt574T/A) promote viral replication by inhibiting the AKT-mTOR autophagy pathway.
Other co-mutations (R142K-nt88C/T, R142K-nt157G/A) reduce viral replication by lessening the inhibition of the AKT-mTOR pathway.

Read full analysis

Keywords:

3D protein 5′UTR AKT-mTOR pathway Autophagy Co-mutation EV71

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