Functional and Physical Interaction between the Arf Activator GBF1 and Hepatitis C Virus NS3 Protein

Lebsir, Nadjet; Goueslain, Lucie; Farhat, Rayan; Callens, Nathalie; Dubuisson, Jean; Jackson, Catherine; Rouillé, Yves

doi:10.1128/jvi.01459-18

Cited by 18 publications

(8 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Here we have not addressed the phosphorylation change in NLRP3 upon HCV infection, but we show that its expression is induced upon HCV infection, that it associates with ASC and importantly that its depletion by silencing prevents Golgi fragmentation. Alternatively, the HCV NS3 protein has been reported to interact with IRGM (Gregoire et al, 2011) and also with GBF1 (Lebsir et al, 2019) and could play a role in the activation of the NLRP3 inflammasome. Whatever the case, since we have shown a direct link between NLRP3 and the IRGM-mediated Golgi fragmentation, our data emphasize the importance of NLRP3 in HCV infection and highlight the role of the ASC/IRGM interaction at the Golgi.…”

Section: Discussionmentioning

confidence: 99%

The inflammasome components NLRP3 and ASC act in concert with IRGM to rearrange the Golgi during viral infections

Daussy

Monard

Guy

et al. 2020

Preprint

View full text Add to dashboard Cite

Hepatitis C virus (HCV) infection triggers Golgi fragmentation through the Golgi-resident protein immunity-related GTPase M (IRGM). Here, we report the role of NLRP3 and ASC, two inflammasome components, in the initial events leading to this fragmentation. We show that ASC resides at the Golgi with IRGM at homeostasis. Upon infection, ASC dissociates from both IRGM and Golgi and associates with HCV-induced NLRP3. NLRP3 silencing inhibits Golgi fragmentation. ASC silencing disrupts the Golgi structure in both control and infected cells and reduces the localization of IRGM at the Golgi.Silencing IRGM cannot totally restore the Golgi structure. These data highlight a role for ASC, upstream of the formation of the inflammasome, in regulating IRGM through its control on the Golgi. A similar mechanism occurs in response to Nigericin or infection with Zika virus (ZIKV). We propose a model for a newly ascribed function of the inflammasome components in Golgi structural remodeling. proteins involved in innate immune response leading to activation of caspase-1 and secretion of the pro-inflammatory cytokines IL-1ß and IL-18. Formation of the NLRP3 inflammasome complex was reported to occur also in hepatocytes, although of non-immune lineage, but in this case without secretion of the pro-inflammatory cytokines IL-1ß and IL-18 (Negash & Gale, 2015). In particular, HCV infection induces the activation of the NLRP3 inflammasome in cultured hepatoma cells that can ultimately lead to the caspase-1-mediated form of programmed cell death called pyroptosis (Kofahi, Taylor, Hirasawa, Grant, & Russell, 2016), Whatever the nature of the cells, activation of the NLRP3 inflammasome requires association of NLRP3 to caspase-1 through ASC (adaptor protein apoptosisassociated speck-like protein containing a CARD). IRGM contains a globular N-terminal GTPase domain and, like ASC, a C-terminal helical CaspaseActivation and Recruitment Domain (CARD). Here we show that IRGM interacts with ASC at the Golgi in homeostasis conditions. Upon HCV infection, NLRP3 is induced and ASC dissociates from both the Golgi and from IRGM. The ability of IRGM to mediate Golgi fragmentation was reduced in the absence of NLRP3 or ASC. Similar molecular events were observed in response to Nigericin, or upon infection of macrophages with Zika virus (ZIKV), thus assigning a newly defined function of the NLRP3 inflammasome components in the regulators of the host intracellular compartments upon infections.

show abstract

Section: Discussionmentioning

confidence: 99%

The inflammasome components NLRP3 and ASC act in concert with IRGM to rearrange the Golgi during viral infections

Daussy

Monard

Guy

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…In the case of influenza (61, 62) and vesicular stomatitis viruses (VSV) (63, 64) the COPI/Arf1 complex seems to play a role in their entry process. Moreover, for VSV as well as for hepatitis C virus (65–67), mouse hepatitis coronavirus (68), chikungunya virus (69), poliovirus (57, 58, 70), coxsackievirus (71) and classical swine fever virus (72), COPI/Arf1 has been shown to be necessary for genome replication as well as for viral protein expression. COPI/Arf1 has also been shown to be critical for viral particle assembly of influenza and the Chandipura virus (62, 73).…”

Section: Discussionmentioning

confidence: 99%

The guanine nucleotide exchange factor GBF1 participates in rotavirus replication

Martínez

Arnoldi

Schraner

et al. 2019

Preprint

View full text Add to dashboard Cite

19Cellular and viral factors participate in the replication cycle of rotavirus. We report that the 20 guanine nucleotide exchange factor GBF1, which activates the small GTPase Arf1 to induce 21 COPI transport processes, is required for rotavirus replication since knocking down GBF1 22 3 IMPORTANCE 40 Rotavirus, a member of the family Reoviridae, is the major cause of severe diarrhea in 41 children and young animals worldwide. Despite the significant advances in the 42 characterization of the biology of this virus, the mechanisms involved in morphogenesis of 43 the virus particle are still poorly understood. In this work, we show that the guanine 44 nucleotide exchange factor GBF1, relevant for the COPI/Arf1-mediated cellular vesicular 45 transport, participates in the replication cycle of the virus, influencing the correct processing 46 of viral glycoproteins VP7 and NSP4, and the assembly of the virus surface proteins VP7 47 59 Rotaviruses, members of the family Reoviridae, are non-enveloped particles formed by three 60 concentric layers of proteins that surround the eleven genome segments of double-stranded 61 RNA (dsRNA). The innermost layer is composed of the core-shell proteinVP2 that encloses 62 the replication intermediates, composed of the RNA dependent RNA polymerase VP1, and 63 the guanylyl-methyl transferase, VP3. The intermediate layer is formed by VP6 that 64 surrounds the VP2 layer to form double-layered particles (DLPs). Finally, the addition of the 65 glycoprotein VP7 and the spike protein VP4 onto the DLPs forms the infectious triple-layered 66 particles (TLPs) (1, 2). 67 The replication of rotavirus occurs in cytoplasmic non-membranous electron-dense 68 inclusions termed viroplasms composed of NSP2, NSP5, VP1, VP2, VP6 and host 69 components (1, 3). The replication and packaging of the viral genome into newly synthesized 70DLPs take place in these inclusions (4), which then bud into the lumen of the endoplasmic 71 reticulum (ER) through membrane sites modified by the presence of NSP4 (5, 6). NSP4 is a 72 transmembrane ER glycoprotein with two N-linked high mannose glycosylated chains (7) 73 that play a crucial role in the last steps of rotavirus assembly. It has been shown that the 74 cytoplasm oriented-terminus of NSP4 associates with VP4 (8), and binds the VP6 on DLPs 75 acting as a receptor for these particles to mediate their budding into the ER (9, 10). Moreover, 76 NSP4 has been shown to also interact with VP7 through its N-terminus oriented to the ER 77 lumen (11, 12). It has been proposed that these interactions drive the incorporation of the 78 outer layer proteins into the transitory lipid envelope that DLPs acquire during ER membrane 79 budding; this envelope is removed in the lumen of the ER by an unknown process in which 80 NSP4 is eliminated while VP4 and VP7 are assembled to produce the final infectious TLPs 81 All rights reserved. No reuse allowed without permission.The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.110...

show abstract

“…In the case of influenza virus (62,63) and vesicular stomatitis virus (VSV) (64,65), the COPI/Arf1 complex seems to play a role in their entry process. Moreover, for VSV as well as for hepatitis C virus (66)(67)(68), mouse hepatitis coronavirus (69), chikungunya virus (70), poliovirus (58,59,71), coxsackievirus (72), and classical swine fever virus (73), COPI/Arf1 has been shown to be necessary for genome replication as well as for viral protein expression. COPI/Arf1 has also been shown to be critical for viral particle assembly of influenza virus and the Chandipura virus (63,74).…”

Section: Discussionmentioning

confidence: 99%

The Guanine Nucleotide Exchange Factor GBF1 Participates in Rotavirus Replication

Martínez

Arnoldi

Schraner

et al. 2019

J Virol

View full text Add to dashboard Cite

Cellular and viral factors participate in the replication cycle of rotavirus. We report that the guanine nucleotide exchange factor GBF1, which activates the small GTPase Arf1 to induce COPI transport processes, is required for rotavirus replication since knocking down GBF1 expression by RNA interference or inhibiting its activity by treatment with brefeldin A (BFA) or Golgicide A (GCA) significantly reduces the yield of infectious viral progeny. This reduction in virus yield was related to a block in virus assembly, since in the presence of either BFA or GCA, the assembly of infectious mature triple-layered virions was significantly prevented and only double-layered particles were detected. We report that the catalytic activity of GBF1, but not the activation of Arf1, is essential for the assembly of the outer capsid of rotavirus. We show that both BFA and GCA, as well as interfering with the synthesis of GBF1, alter the electrophoretic mobility of glycoproteins VP7 and NSP4 and block the trimerization of the virus surface protein VP7, a step required for its incorporation into virus particles. Although a posttranslational modification of VP7 (other than glycosylation) could be related to the lack of trimerization, we found that NSP4 might also be involved in this process, since knocking down its expression reduces VP7 trimerization. In support, recombinant VP7 protein overexpressed in transfected cells formed trimers only when cotransfected with NSP4. IMPORTANCE Rotavirus, a member of the family Reoviridae, is the major cause of severe diarrhea in children and young animals worldwide. Despite significant advances in the characterization of the biology of this virus, the mechanisms involved in morphogenesis of the virus particle are still poorly understood. In this work, we show that the guanine nucleotide exchange factor GBF1, relevant for COPI/Arf1mediated cellular vesicular transport, participates in the replication cycle of the virus, influencing the correct processing of viral glycoproteins VP7 and NSP4 and the assembly of the virus surface proteins VP7 and VP4. KEYWORDS GBF1 nucleotide exchange factor, morphogenesis, rotavirus R otaviruses, members of the family Reoviridae, are nonenveloped particles formed by three concentric layers of proteins that surround the 11 genome segments of double-stranded RNA (dsRNA). The innermost layer is composed of the core shell protein VP2, which encloses the replication intermediates, composed of the RNAdependent RNA polymerase VP1 and the guanylyl-methyl transferase VP3. The intermediate layer is formed by VP6, which surrounds the VP2 layer to form double-layered particles (DLPs). Finally, the addition of the glycoprotein VP7 and the spike protein VP4 onto the DLPs forms the infectious triple-layered particles (TLPs) (1, 2). Downloaded fromFIG 2 BFA and GCA inhibit rotavirus replication at a postentry step. (A) MA104 cells were infected with RRV (MOI ϭ 5) at 37°C for 1 h. Unbound virus was removed, and 0.5 g/ml of BFA or 10 M GCA was added at the indicated times postinf...

show abstract

Functional and Physical Interaction between the Arf Activator GBF1 and Hepatitis C Virus NS3 Protein

Cited by 18 publications

References 63 publications

The inflammasome components NLRP3 and ASC act in concert with IRGM to rearrange the Golgi during viral infections

The inflammasome components NLRP3 and ASC act in concert with IRGM to rearrange the Golgi during viral infections

The guanine nucleotide exchange factor GBF1 participates in rotavirus replication

The Guanine Nucleotide Exchange Factor GBF1 Participates in Rotavirus Replication

Contact Info

Product

Resources

About