The Golgi Protein ACBD3, an Interactor for Poliovirus Protein 3A, Modulates Poliovirus Replication

Téoulé, François; Brisac, Cynthia; Pelletier, Isabelle; Vidalain, Pierre‐Olivier; Jégouic, Sophie; Mirabelli, Carmen; Bessaud, Maël; Combelas, Nicolas; Autret, Arnaud; Tangy, Frédéric; Delpeyroux, Françis; Blondel, B

doi:10.1128/jvi.00304-13

Cited by 45 publications

(42 citation statements)

References 56 publications

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“…To investigate the importance of GBF1 activity for PI4KIII␤ recruitment, we utilized an in vitro system with HeLa S10 cell extracts (21). RNA coding for CVB3 3A was translated in these cell extracts in the absence or presence of brefeldin A (BFA), a well-known inhibitor of GBF1 (34). Subsequently, the membranes were isolated by centrifugation and the membrane-associated proteins were analyzed by Western blotting.…”

Section: Resultsmentioning

confidence: 99%

Recruitment of PI4KIIIβ to Coxsackievirus B3 Replication Organelles Is Independent of ACBD3, GBF1, and Arf1

Dorobantu

Schaar

Ford

et al. 2014

J Virol

105

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Members of the Enterovirus (poliovirus [PV], coxsackieviruses, and human rhinoviruses) and Kobuvirus (Aichi virus) genera in the Picornaviridae family rely on PI4KIII␤ (phosphatidylinositol-4-kinase III␤) for efficient replication. The small membraneanchored enteroviral protein 3A recruits PI4KIII␤ to replication organelles, yet the underlying mechanism has remained elusive. Recently, it was shown that kobuviruses recruit PI4KIII␤ through interaction with ACBD3 (acyl coenzyme A [acyl-CoA]-binding protein domain 3), a novel interaction partner of PI4KIII␤. Therefore, we investigated a possible role for ACBD3 in recruiting PI4KIII␤ to enterovirus replication organelles. Although ACBD3 interacted directly with coxsackievirus B3 (CVB3) 3A, its depletion from cells by RNA interference did not affect PI4KIII␤ recruitment to replication organelles and did not impair CVB3 RNA replication. Enterovirus 3A was previously also proposed to recruit PI4KIII␤ via GBF1/Arf1, based on the known interaction of 3A with GBF1, an important regulator of secretory pathway transport and a guanine nucleotide exchange factor (GEF) of Arf1. However, our results demonstrate that inhibition of GBF1 or Arf1 either by pharmacological inhibition or depletion with small interfering RNA (siRNA) treatment did not affect the ability of 3A to recruit PI4KIII␤. Furthermore, we show that a 3A mutant that no longer binds GBF1 was capable of recruiting PI4KIII␤, even in ACBD3-depleted cells. Together, our findings indicate that unlike originally envisaged, coxsackievirus recruits PI4KIII␤ to replication organelles independently of ACBD3 and GBF1/Arf1. IMPORTANCEA hallmark of enteroviral infection is the generation of new membranous structures to support viral RNA replication. The functionality of these "replication organelles" depends on the concerted actions of both viral nonstructural proteins and co-opted host factors. It is thus essential to understand how these structures are formed and which cellular components are key players in this process. GBF1/Arf1 and ACBD3 have been proposed to contribute to the recruitment of the essential lipid-modifying enzyme PI4KIII␤ to enterovirus replication organelles. Here we show that the enterovirus CVB3 recruits PI4KIII␤ by a mechanism independent of both GBF1/Arf1 and ACBD3. This study shows that the strategy employed by coxsackievirus to recruit PI4KIII␤ to replication organelles is far more complex than initially anticipated.

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Section: Resultsmentioning

confidence: 99%

Recruitment of PI4KIIIβ to Coxsackievirus B3 Replication Organelles Is Independent of ACBD3, GBF1, and Arf1

Dorobantu

Schaar

Ford

et al. 2014

J Virol

105

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“…CVB3 3A and PV 3A both interact with the N terminus of GBF1 (7,8) as well as ACBD3 (9). However, we and others recently showed that the PI4KIII␤ recruitment by 3A of PV and CVB3 seems to occur independently of both GBF1/Arf1 and ACBD3 (9,10). We studied this by individually expressing mutant 3A proteins of CVB3 that no longer interact with GBF1 (9).…”

mentioning

confidence: 99%

GBF1- and ACBD3-Independent Recruitment of PI4KIIIβ to Replication Sites by Rhinovirus 3A Proteins

Dorobantu

Ford-Siltz

Sittig

et al. 2015

J Virol

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c PI4KIII␤ recruitment to Golgi membranes relies on GBF1/Arf and ACBD3. Enteroviruses such as poliovirus and coxsackievirus recruit PI4KIII␤ to their replication sites via their 3A proteins. Here, we show that human rhinovirus (HRV) 3A also recruited PI4KIII␤ to replication sites. Unlike other enterovirus 3A proteins, HRV 3A failed to bind GBF1. Although HRV 3A was previously shown to interact with ACBD3, our data suggest that PI4KIII␤ recruitment occurred independently of both GBF1 and ACBD3. E nteroviruses (family Picornaviridae), such as poliovirus (PV) and coxsackievirus B3 (CVB3), rely on host factor phosphatidylinositol-4-kinase III␤ (PI4KIII␤) for genome replication (1, 2). The recruitment of PI4KIII␤ to the replication sites is mediated by the 3A viral nonstructural protein (1). PI4KIII␤ is normally recruited to the Golgi membranes by the small GTPase Arf1 (3) and the Arf1 activator guanine nucleotide exchange factor (GEF) GBF1 (4). ACBD3 (acyl-coenzyme A [CoA]-binding protein domain 3) also interacts with PI4KIII␤ to recruit it to the Golgi membranes (5, 6). CVB3 3A and PV 3A both interact with the N terminus of GBF1 (7, 8) as well as ACBD3 (9). However, we and others recently showed that the PI4KIII␤ recruitment by 3A of PV and CVB3 seems to occur independently of both GBF1/Arf1 and ACBD3 (9, 10). We studied this by individually expressing mutant 3A proteins of CVB3 that no longer interact with GBF1 (9). Unfortunately, these 3A mutations render CVB3 unviable (F. J. M. van Kuppeveld, unpublished data); thus, we have not yet been able to study the role of GBF1 in PI4KIII␤ recruitment in infected cells. Yeast two-hybrid analysis suggests that the 3A proteins of human rhinoviruses (HRV), which also belong to the Enterovirus genus, do not interact with the N terminus of GBF1 (8). In this study, we set out to investigate the consequences of the inability of HRV 3A proteins to interact with GBF1, focusing on the possible role of GBF1 and ACBD3 in PI4KIII␤ recruitment and virus replication.First, we studied the interaction between the N terminus of GBF1 and the 3A proteins of HRV2 and HRV14, which are members of group A and B rhinoviruses, respectively, in a mammalian environment, employing the mammalian two-hybrid (M2H) assay (Promega) as described previously (9). We used the 3As of CVB3, PV, and mengovirus, belonging to the Cardiovirus genus of the Picornaviridae, as controls. Expression of all 3A proteins was verified by Western blot analysis (Fig. 1A, lower half). Importantly, all enterovirus 3A proteins were previously tested and validated to be competent in interacting with other proteins in the M2H assay (9). Both CVB3 3A and PV 3A interacted with the N terminus of GBF1, while no interaction was detected for mengovirus 3A (Fig. 1A, upper panel). HRV2 3A did not bind the N terminus of GBF1, while HRV14 3A interacted to only a limited extent. We considered the possibility that the HRV 3A proteins bind to the C terminus of GBF1. Therefore, we studied whether 3A expression induced recruitment of full-length...

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“…This pathway transiently protects cells against PV-induced cell death and may provide the virus with sufficient time to replicate. We previously showed that the non-structural 3A protein of PV interacts with CREB3 (Teoule et al, 2013). However, the role of the 3A protein in this process remains to be investigated.…”

Section: Herpmentioning

confidence: 99%

“…Herp is under the transcriptional control of cAMP response element-binding protein 3 (CREB3), a member of the ER membrane-bound CREB/ATF (activating transcription factor) family of proteins, which is rapidly activated in response to cellular stress signals via the Golgi regulated intramembrane proteolysis (RIP) pathway (Asada et al, 2011;Chan et al, 2011;Raggo et al, 2002). We had also identified CREB as interacting with the PV nonstructural protein 3A (Teoule et al, 2013), a multifunction protein involved in PV replication, both directly and indirectly, via massive remodeling of host intracellular membranes (Belov & van Kuppeveld, 2012;Paul & Wimmer, 2015;van Kuppeveld et al, 2010). We therefore hypothesized that PVinduced Ca 2+ release from the ER and apoptosis might be modulated by the CREB3/Herp signalling pathway.…”

mentioning

confidence: 99%

“…Cells were also treated with thapsigargin (TG, 1 µM, Sigma-Aldrich T9033), a powerful Herp protein inducer (Kokame et al, 2000;Thastrup et al, 1990), as a positive control. Whole-cell extracts were collected at the times indicated and analysed by Western blotting as previously described (Teoule et al, 2013), with an anti-Herp antibody (19-Y, sc-100721, Santacruz) (Fig. 1a).…”

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confidence: 99%

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The CREB3-Herp signalling module limits the cytosolic calcium concentration increase and apoptosis induced by poliovirus

Mirabelli

Pelletier

Téoulé

et al. 2016

Journal of General Virology

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The Golgi Protein ACBD3, an Interactor for Poliovirus Protein 3A, Modulates Poliovirus Replication

Cited by 45 publications

References 56 publications

Recruitment of PI4KIIIβ to Coxsackievirus B3 Replication Organelles Is Independent of ACBD3, GBF1, and Arf1

Recruitment of PI4KIIIβ to Coxsackievirus B3 Replication Organelles Is Independent of ACBD3, GBF1, and Arf1

GBF1- and ACBD3-Independent Recruitment of PI4KIIIβ to Replication Sites by Rhinovirus 3A Proteins

The CREB3-Herp signalling module limits the cytosolic calcium concentration increase and apoptosis induced by poliovirus

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