A Dominant-Negative Mutant of rab5 Inhibits Infection of Cells by Foot-and-Mouth Disease Virus: Implications for Virus Entry

ABSTRACT Foot-and-mouth disease virus (FMDV) can use a number of different integrins (αvβ1, αvβ3, αvβ6, and αvβ8) as receptors to initiate infection. Infection mediated by αvβ6 is known to occur by clathrin-mediated endocytosis and is dependent on the acidic pH within endosomes. On internalization, virus is detected rapidly in early endosomes (EE) and subsequently in perinuclear recycling endosomes (PNRE), but not in late endosomal compartments. Due to the extreme sensitivity of FMDV to acidic pH, it is thought that EE can provide a pH low enough for infection to occur; however, definitive proof that infection takes place from within these compartments is still lacking. Here we have investigated the intracellular transport steps required for FMDV infection of IBRS-2 cells, which express αvβ8 as their FMDV receptor. These experiments confirmed that FMDV infection mediated by αvβ8 is also dependent on clathrin-mediate endocytosis and an acidic pH within endosomes. Also, the effect on FMDV infection of dominant-negative (DN) mutants of cellular rab proteins that regulate endosomal traffic was examined. Expression of DN rab5 reduced the number of FMDV-infected cells by 80%, while expression of DN rab4 or DN rab7 had virtually no effect on infection. Expression of DN rab11 inhibited infection by FMDV, albeit to a small extent (∼35%). These results demonstrate that FMDV infection takes place predominantly from within EE and does not require virus trafficking to the late endosomal compartments. However, our results suggest that infection may not be exclusive to EE and that a small amount of infection could occur from within PNRE.

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A role for endoplasmic reticulum exit sites in foot-and-mouth disease virus infection

Journal of General Virology ◽

10.1099/vir.0.055442-0 ◽

2013 ◽

Vol 94 (12) ◽

pp. 2636-2646 ◽

Cited By ~ 19

Author(s):

Rebecca Midgley ◽

Katy Moffat ◽

Stephen Berryman ◽

Philippa Hawes ◽

Jennifer Simpson ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Disease Virus ◽

Secretory Pathway ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Rab Proteins ◽

Fmdv Infection ◽

Early Secretory Pathway ◽

Mouth Disease Virus ◽

Foot And Mouth

Picornaviruses replicate their genomes in association with cellular membranes. While enteroviruses are believed to utilize membranes of the early secretory pathway, the origin of the membranes used by foot-and-mouth disease virus (FMDV) for replication are unknown. Secretory-vesicle traffic through the early secretory pathway is mediated by the sequential acquisition of two distinct membrane coat complexes, COPII and COPI, and requires the coordinated actions of Sar1, Arf1 and Rab proteins. Sar1 is essential for generating COPII vesicles at endoplasmic reticulum (ER) exit sites (ERESs), while Arf1 and Rab1 are required for subsequent vesicle transport by COPI vesicles. In the present study, we have provided evidence that FMDV requires pre-Golgi membranes of the early secretory pathway for infection. Small interfering RNA depletion of Sar1 or expression of a dominant-negative (DN) mutant of Sar1a inhibited FMDV infection. In contrast, a dominant-active mutant of Sar1a, which allowed COPII vesicle formation but inhibited the secretory pathway by stabilizing COPII coats, caused major disruption to the ER–Golgi intermediate compartment (ERGIC) but did not inhibit infection. Treatment of cells with brefeldin A, or expression of DN mutants of Arf1 and Rab1a, disrupted the Golgi and enhanced FMDV infection. These results show that reagents that block the early secretory pathway at ERESs have an inhibitory effect on FMDV infection, while reagents that block the early secretory pathway immediately after ER exit but before the ERGIC and Golgi make infection more favourable. Together, these observations argue for a role for Sar1 in FMDV infection and that initial virus replication takes place on membranes that are formed at ERESs.

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Proteomics Analysis of Porcine Serum Proteins by LC-MS/MS after Foot-and-Mouth Disease Virus (FMDV) Infection

Journal of Veterinary Medical Science ◽

10.1292/jvms.11-0019 ◽

2011 ◽

Vol 73 (12) ◽

pp. 1569-1572 ◽

Cited By ~ 7

Author(s):

Yongjie LIU ◽

Keshan ZHANG ◽

Haixue ZHENG ◽

Youjun SHANG ◽

Jianhong GUO ◽

...

Keyword(s):

Disease Virus ◽

Serum Proteins ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Proteomics Analysis ◽

Porcine Serum ◽

Fmdv Infection ◽

Mouth Disease Virus ◽

Foot And Mouth

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Establishment of persistent foot-and-mouth disease virus (FMDV) infection in MDBK cells

Archives of Virology ◽

10.1007/s00705-015-2526-8 ◽

2015 ◽

Vol 160 (10) ◽

pp. 2503-2516 ◽

Cited By ~ 7

Author(s):

Lela Kopliku ◽

Anthony Relmy ◽

Aurore Romey ◽

Kamila Gorna ◽

Stephan Zientara ◽

...

Keyword(s):

Disease Virus ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Fmdv Infection ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Mdbk Cells

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The Carrier Conundrum; A Review of Recent Advances and Persistent Gaps Regarding the Carrier State of Foot-and-Mouth Disease Virus

Pathogens ◽

10.3390/pathogens9030167 ◽

2020 ◽

Vol 9 (3) ◽

pp. 167 ◽

Cited By ~ 2

Author(s):

Carolina Stenfeldt ◽

Jonathan Arzt

Keyword(s):

Disease Virus ◽

Foot And Mouth Disease ◽

Field Studies ◽

Carrier State ◽

Mouth Disease ◽

Experimental Investigations ◽

Apparent Lack ◽

Fmdv Infection ◽

Mouth Disease Virus ◽

Foot And Mouth

The existence of a prolonged, subclinical phase of foot-and-mouth disease virus (FMDV) infection in cattle was first recognized in the 1950s. Since then, the FMDV carrier state has been a subject of controversy amongst scientists and policymakers. A fundamental conundrum remains in the discordance between the detection of infectious FMDV in carriers and the apparent lack of contagiousness to in-contact animals. Although substantial progress has been made in elucidating the causal mechanisms of persistent FMDV infection, there are still critical knowledge gaps that need to be addressed in order to elucidate, predict, prevent, and model the risks associated with the carrier state. This is further complicated by the occurrence of a distinct form of neoteric subclinical infection, which is indistinguishable from the carrier state in field scenarios, but may have substantially different epidemiological properties. This review summarizes the current state of knowledge of the FMDV carrier state and identifies specific areas of research in need of further attention. Findings from experimental investigations of FMDV pathogenesis are discussed in relation to experience gained from field studies of foot-and-mouth disease.

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Sec62 Regulates Endoplasmic Reticulum Stress and Autophagy Balance to Affect Foot-and-Mouth Disease Virus Replication

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.707107 ◽

2021 ◽

Vol 11 ◽

Author(s):

Jin’en Wu ◽

Zhihui Zhang ◽

Zhidong Teng ◽

Sahibzada Waheed Abdullah ◽

Shiqi Sun ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Er Stress ◽

Disease Virus ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Jnk Pathway ◽

Fmdv Infection ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Fmdv Replication

Endoplasmic reticulum (ER) stress-induced autophagy is closely associated with viral infection and propagation. However, the intrinsic link between ER stress, autophagy, and viral replication during foot-and-mouth disease virus (FMDV) infection is not fully elucidated. Our previous studies demonstrated that FMDV infection activated the ER stress-associated UPR of the PERK-eIF2a and ATF6 signaling pathway, whereas the IRE1a signaling was suppressed. We found that the activated-ATF6 pathway participated in FMDV-induced autophagy and FMDV replication, while the IRE1α pathway only affected FMDV replication. Further studies indicated that Sec62 was greatly reduced in the later stages of FMDV infection and blocked the activation of the autophagy-related IRE1α-JNK pathway. Moreover, it was also found that Sec62 promoted IRE1a phosphorylation and negatively regulated FMDV proliferation. Importantly, Sec62 may interact with LC3 to regulate ER stress and autophagy balance and eventually contribute to FMDV clearance via fusing with lysosomes. Altogether, these results suggest that Sec62 is a critical molecule in maintaining and recovering ER homeostasis by activating the IRE1α-JNK pathway and delivering autophagosome into the lysosome, thus providing new insights on FMDV-host interactions and novel antiviral therapies.

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An infectious recombinant foot-and-mouth disease virus expressing a fluorescent marker protein

Journal of General Virology ◽

10.1099/vir.0.052308-0 ◽

2013 ◽

Vol 94 (7) ◽

pp. 1517-1527 ◽

Cited By ~ 20

Author(s):

Julian Seago ◽

Nicholas Juleff ◽

Katy Moffat ◽

Stephen Berryman ◽

John M. Christie ◽

...

Keyword(s):

Disease Virus ◽

Reverse Genetics ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Marker Protein ◽

Fluorescent Marker ◽

Fmdv Infection ◽

Animal Pathogens ◽

Mouth Disease Virus ◽

Foot And Mouth

Foot-and-mouth disease virus (FMDV) is one of the most extensively studied animal pathogens because it remains a major threat to livestock economies worldwide. However, the dynamics of FMDV infection are still poorly understood. The application of reverse genetics provides the opportunity to generate molecular tools to further dissect the FMDV life cycle. Here, we have used reverse genetics to determine the capsid packaging limitations for a selected insertion site in the FMDV genome. We show that exogenous RNA up to a defined length can be stably introduced into the FMDV genome, whereas larger insertions are excised by recombination events. This led us to construct a recombinant FMDV expressing the fluorescent marker protein, termed iLOV. Characterization of infectious iLOV-FMDV showed the virus has a plaque morphology and rate of growth similar to the parental virus. In addition, we show that cells infected with iLOV-FMDV are easily differentiated by flow cytometry using the inherent fluorescence of iLOV and that cells infected with iLOV-FMDV can be monitored in real-time with fluorescence microscopy. iLOV-FMDV therefore offers a unique tool to characterize FMDV infection in vitro, and its applications for in vivo studies are discussed.

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Cell mediated innate responses of cattle and swine are diverse during foot-and-mouth disease virus (FMDV) infection: A unique landscape of innate immunity

Immunology Letters ◽

10.1016/j.imlet.2013.05.007 ◽

2013 ◽

Vol 152 (2) ◽

pp. 135-143 ◽

Cited By ~ 25

Author(s):

Felix N. Toka ◽

William T. Golde

Keyword(s):

Innate Immunity ◽

Disease Virus ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Fmdv Infection ◽

Mouth Disease Virus ◽

Foot And Mouth

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Discriminating Foot-and-Mouth Disease Virus-Infected and Vaccinated Animals by Use of β-Galactosidase Allosteric Biosensors

Clinical and Vaccine Immunology ◽

10.1128/cvi.00139-09 ◽

2009 ◽

Vol 16 (8) ◽

pp. 1228-1235 ◽

Cited By ~ 6

Author(s):

M. Teresa Sánchez-Aparicio ◽

María Flora Rosas ◽

Rosa Maria Ferraz ◽

Laura Delgui ◽

Juan J. Veloso ◽

...

Keyword(s):

Disease Virus ◽

Nonstructural Protein ◽

Polyclonal Antibodies ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Reaction Conditions ◽

Fmdv Infection ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Cattle And Sheep

ABSTRACT Recombinant β-galactosidases accommodating one or two different peptides from the foot-and-mouth disease virus (FMDV) nonstructural protein 3B per enzyme monomer showed a drastic enzymatic activity reduction, which mainly affected proteins with double insertions. Recombinant β-galactosidases were enzymatically reactivated by 3B-specific murine monoclonal and rabbit polyclonal antibodies. Interestingly, these recombinant β-galactosidases, particularly those including one copy of each of the two 3B sequences, were efficiently reactivated by sera from infected pigs. We found reaction conditions that allowed differentiation between sera of FMDV-infected pigs, cattle, and sheep and those of naïve and conventionally vaccinated animals. These FMDV infection-specific biosensors can provide an effective and versatile alternative for the serological distinction of FMDV-infected animals.

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Internalization of Swine Vesicular Disease Virus into Cultured Cells: a Comparative Study with Foot-and-Mouth Disease Virus

Journal of Virology ◽

10.1128/jvi.02436-08 ◽

2009 ◽

Vol 83 (9) ◽

pp. 4216-4226 ◽

Cited By ~ 10

Author(s):

Miguel A. Martín-Acebes ◽

Mónica González-Magaldi ◽

Angela Vázquez-Calvo ◽

Rosario Armas-Portela ◽

Francisco Sobrino

Keyword(s):

Disease Virus ◽

Cultured Cells ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Coated Pits ◽

Early Endosomes ◽

Swine Vesicular Disease Virus ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Vesicular Disease

ABSTRACT We performed a comparative analysis of the internalization mechanisms used by three viruses causing important vesicular diseases in animals. Swine vesicular disease virus (SVDV) internalization was inhibited by treatments that affected clathrin-mediated endocytosis and required traffic through an endosomal compartment. SVDV particles were found in clathrin-coated pits by electron microscopy and colocalized with markers of early endosomes by confocal microscopy. SVDV infectivity was significantly inhibited by drugs that raised endosomal pH. When compared to foot-and-mouth disease virus (FMDV), which uses clathrin-mediated endocytosis, the early step of SVDV was dependent on the integrity of microtubules. SVDV-productive endocytosis was more sensitive to plasma membrane cholesterol extraction than that of FMDV, and differential cell signaling requirements for virus infection were also found. Vesicular stomatitis virus, a model virus internalized by clathrin-mediated endocytosis, was included as a control of drug treatments. These results suggest that different clathrin-mediated routes are responsible for the internalization of these viruses.

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