Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014 Jun 9;9(6):e99768.
doi: 10.1371/journal.pone.0099768. eCollection 2014.

Class I ADP-ribosylation factors are involved in enterovirus 71 replication

Jianmin Wang  1 , Jiang Du  1 , Qi Jin  1
Affiliations

Class I ADP-ribosylation factors are involved in enterovirus 71 replication

Jianmin Wang et al. PLoS One. .

Abstract

Enterovirus 71 is one of the major causative agents of hand, foot, and mouth disease in infants and children. Replication of enterovirus 71 depends on host cellular factors. The viral replication complex is formed in novel, cytoplasmic, vesicular compartments. It has not been elucidated which cellular pathways are hijacked by the virus to create these vesicles. Here, we investigated whether proteins associated with the cellular secretory pathway were involved in enterovirus 71 replication. We used a loss-of-function assay, based on small interfering RNA. We showed that enterovirus 71 RNA replication was dependent on the activity of Class I ADP-ribosylation factors. Simultaneous depletion of ADP-ribosylation factors 1 and 3, but not three others, inhibited viral replication in cells. We also demonstrated with various techniques that the brefeldin-A-sensitive guanidine nucleotide exchange factor, GBF1, was critically important for enterovirus 71 replication. Our results suggested that enterovirus 71 replication depended on GBF1-mediated activation of Class I ADP-ribosylation factors. These results revealed a connection between enterovirus 71 replication and the cellular secretory pathway; this pathway may represent a novel target for antiviral therapies.

PubMed Disclaimer

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Class I Arfs are upregulated and activated in EV71 infections.
(A) Transcripts for both Arf1 and Arf3 are upregulated in EV71-infected cells (P<0.05). (B) Expression levels of Class I Arf proteins are upregulated with EV71 infection. (C) Class I Arfs are activated upon EV71 infection.
Figure 2
Figure 2. Knockdown of a single Arf does not affect EV71 replication in cells.
(A) Individual Arf isoforms were effectively knocked-down with siRNAs (P<0.05). (B) Knockdown of a single Arf did not affect EV71 replication in cells (P>0.05).
Figure 3
Figure 3. Double knockdown of Arf1 and Arf3 inhibits EV71 replication in RD cells.
Of all the possible Arf pairs (omitting Arf2), only the Arf1+Arf3 double knockdown inhibited EV71 replication (P<0.05).
Figure 4
Figure 4. Overexpression of Arf proteins could not rescue EV71 replication from BFA exposure.
(A) All the Arf proteins tested are expressed at high levels in RD cells. (B) Overexpression of Arf proteins did not rescue viral replication under BFA exposure (P>0.05).
Figure 5
Figure 5. GBF1 is required for EV71 replication.
(A) Targeted siRNAs could effectively knockdown the expression of each GEF (P<0.05). (B) Knockdown of GBF1, but not BIG1 or BIG2, inhibited EV71 replication in cells (P<0.05). (C) GBF1-EGFP is overexpressed in RD cells by transfection. (D) Overexpression of GBF1 rescued viral replication from BFA exposure (P<0.05).
Figure 6
Figure 6. GBF1 interacts with viral 3A protein.
(A) Immunoprecipitation was conducted with Protein G agarose plus anti-FLAG antibody. Western blots were probed with specific antibodies, as indicated. (B) Immunoprecipitation was conducted with Protein G agarose plus anti-GFP antibody. Western blots were probed with specific antibodies, as indicated.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Liao HT, Hung KL (2001) Neurologic involvement in an outbreak of enterovirus 71 infection: a hospital-based study. Acta Paediatr Taiwan 42: 27–32. - PubMed
    1. Li L, He Y, Yang H, Zhu J, Xu X, et al. (2005) Genetic characteristics of human enterovirus 71 and coxsackievirus A16 circulating from 1999 to 2004 in Shenzhen, People's Republic of China. J Clin Microbiol 43: 3835–3839. - PMC - PubMed
    1. Buck KW (1996) Comparison of the replication of positive-stranded RNA viruses of plants and animals. Adv Virus Res 47: 159–251. - PMC - PubMed
    1. Bienz K, Egger D, Pfister T, Troxler M (1992) Structural and functional characterization of the poliovirus replication complex. J Virol 66: 2740. - PMC - PubMed
    1. Bienz K, Egger D, Pfister T (1994) Characteristics of the poliovirus replication complex. Arch Virol Suppl 9: 147. - PubMed

Publication types

MeSH terms

Grants and funding

This work was supported by the National Natural Science Foundation of China (No. 31300154) and National Science and Technology Major Project of China (Project No. 2011ZX10004-001 and No. 2013ZX10004-101). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

LinkOut - more resources