ITCH E3 Ubiquitin Ligase Interacts with Ebola Virus VP40 To Regulate Budding

Han, Ziying; Sagum, Cari A.; Bedford, Mark T.; Sidhu, Sachdev S.; Sudol, Marius; Harty, Ronald N.

doi:10.1128/jvi.01078-16

Cited by 62 publications

(66 citation statements)

References 73 publications

(85 reference statements)

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“…The positive effect of host E3 ligase WWP1 on eVP40 VLP budding reported here further highlights the probudding function associated with the host ubiquitination process (1,18,21,65). In contrast, we along with others identified a counteracting, "anti-budding" role for the interferon-induced, ubiquitin-like host protein ISG15 and the process of ISGylation (16,17,66).…”

Section: Discussionmentioning

confidence: 38%

“…We found that the WT peptide interacted with all four of the tandem WW domains within host proteins WWP1 and WWP2, as well as with Itch, as we described previously (Fig. 1A, block C) (21). Here, we will characterize further the physical and functional interactions between eVP40 and WWP1.…”

Section: Resultsmentioning

confidence: 81%

“…The eVP40 PPXY core motif recruits host proteins by interacting specifically with one or more of their modular WW domains (39)(40)(41)(42)(43)(44)(45)(46)(47). Indeed, there is an integral degree of specificity in these virus-host interactions as the eVP40 PPXY motif interacts physically and functionally with a highly select subset of WW-domain-bearing host proteins (21,48,49). Thus, it is of interest to identify and validate the repertoire of WW-domain-containing host protein interactors that may influence or regulate eVP40-mediated egress.…”

mentioning

confidence: 99%

“…To facilitate the efficient release (pinching off) of VLPs, EBOV VP40 (eVP40) recruits multiple host proteins via its late (L) budding domains (PPXY and/or PTAP) during the final stages of virion assembly and egress (1,7,(9)(10)(11)(12)(13)(14)(15). For example, the eVP40 PPXY L-domain motif interacts with HECT family E3 ubiquitin (Ub) ligases Nedd4 (1,6,13,(16)(17)(18)(19)(20) and Itch (21), as well as the cytoskeletal remodeling protein IQGAP1 (12). In general, these virus-host interactions are advantageous for efficient virus production (1,14,15,18,19,(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38).…”

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confidence: 99%

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Ubiquitin Ligase WWP1 Interacts with Ebola Virus VP40 To Regulate Egress

Han

Sagum

Takizawa

et al. 2017

J Virol

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Ebola virus (EBOV) is a member of the Filoviridae family and the cause of hemorrhagic fever outbreaks. The EBOV VP40 (eVP40) matrix protein is the main driving force for virion assembly and budding. Indeed, expression of eVP40 alone in mammalian cells results in the formation and budding of virus-like particles (VLPs) which mimic the budding process and morphology of authentic, infectious EBOV. To complete the budding process, eVP40 utilizes its PPXY L-domain motif to recruit a specific subset of host proteins containing one or more modular WW domains that then function to facilitate efficient production and release of eVP40 VLPs. In this report, we identified additional host WW-domain interactors by screening for potential interactions between mammalian proteins possessing one or more WW domains and WT or PPXY mutant peptides of eVP40. We identified the HECT family E3 ubiquitin ligase WWP1 and all four of its WW domains as strong interactors with the PPXY motif of eVP40. The eVP40-WWP1 interaction was confirmed by both peptide pulldown and coimmunoprecipitation assays, which also demonstrated that modular WW domain 1 of WWP1 was most critical for binding to eVP40. Importantly, the eVP40-WWP1 interaction was found to be biologically relevant for VLP budding since (i) small interfering RNA (siRNA) knockdown of endogenous WWP1 resulted in inhibition of eVP40 VLP egress, (ii) coexpression of WWP1 and eVP40 resulted in ubiquitination of eVP40 and a subsequent increase in eVP40 VLP egress, and (iii) an enzymatically inactive mutant of WWP1 (C890A) did not ubiquitinate eVP40 or enhance eVP40 VLP egress. Last, our data show that ubiquitination of eVP40 by WWP1 enhances egress of VLPs and concomitantly decreases cellular levels of highermolecular-weight oligomers of eVP40. In sum, these findings contribute to our fundamental understanding of the functional interplay between host E3 ligases, ubiquitination, and regulation of EBOV VP40-mediated egress.IMPORTANCE Ebola virus (EBOV) is a high-priority, emerging human pathogen that can cause severe outbreaks of hemorrhagic fever with high mortality rates. As there are currently no approved vaccines or treatments for EBOV, a better understanding of the biology and functions of EBOV-host interactions that promote or inhibit viral budding is warranted. Here, we describe a physical and functional interaction between EBOV VP40 (eVP40) and WWP1, a host E3 ubiquitin ligase that ubiquitinates VP40 and regulates VLP egress. This viral PPXY-host WW domain-mediated interaction represents a potential new target for host-oriented inhibitors of EBOV egress.KEYWORDS E3 ubiquitin ligase, L-domain, PPXY, VLPs, VP40, WW domain, WWP1, budding, Ebola virus

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Section: Discussionmentioning

confidence: 38%

Section: Resultsmentioning

confidence: 81%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Ubiquitin Ligase WWP1 Interacts with Ebola Virus VP40 To Regulate Egress

Han

Sagum

Takizawa

et al. 2017

J Virol

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“…In the case of Ebola infection, EBOV VP35 protein induces the SUMOylation of IRF7 to disrupt antiviral responses12. In addition, ubiquitin itself is thought to be exploited by EBOV to facilitate efficient virus egress31314. However, usurpation of the SUMOylation system by EBOV to regulate its own proteins has not been reported so far.…”

mentioning

confidence: 99%

Regulation of Ebola virus VP40 matrix protein by SUMO

Baz-Martínez

Motiam

Ruibal

et al. 2016

Sci Rep

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The matrix protein of Ebola virus (EBOV) VP40 regulates viral budding, nucleocapsid recruitment, virus structure and stability, viral genome replication and transcription, and has an intrinsic ability to form virus-like particles. The elucidation of the regulation of VP40 functions is essential to identify mechanisms to inhibit viral replication and spread. Post-translational modifications of proteins with ubiquitin-like family members are common mechanisms for the regulation of host and virus multifunctional proteins. Thus far, no SUMOylation of VP40 has been described. Here we demonstrate that VP40 is modified by SUMO and that SUMO is included into the viral like particles (VLPs). We demonstrate that lysine residue 326 in VP40 is involved in SUMOylation, and by analyzing a mutant in this residue we show that SUMO conjugation regulates the stability of VP40 and the incorporation of SUMO into the VLPs. Our study indicates for the first time, to the best of our knowledge, that EBOV hijacks the cellular SUMOylation system in order to modify its own proteins. Modulation of the VP40-SUMO interaction may represent a novel target for the therapy of Ebola virus infection.

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Host Factors Involved in Ebola Virus Replication

Rasmussen

2017

Roles of Host Gene and Non-Coding RNA Expression in Virus Infection

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Ebola virus (EBOV) is a highly pathogenic emerging virus that represents a serious threat to global public health and a major priority for biodefense. The 2014 West African outbreak demonstrated the potential of EBOV to cause an epidemic affecting thousands of people. The severity of disease and high case fatality rate of EBOV is largely due to the host response elicited by the virus. EBOV infection hijacks a number of host pathways to carry out replication and stimulate potent inflammatory responses, while simultaneously subverting the host antiviral immune response. Together, these events trigger a complex, systemic, often lethal febrile disease characterized by high levels of inflammatory cytokines, acute hepatitis and liver dysfunction, immune antagonism, gastrointestinal distress, and, in some cases, hemorrhage caused by coagulopathy and vascular leakage. This review presents current knowledge about the particular host responses induced and disrupted by EBOV infection and how these contribute to virus replication, immune evasion, pathogenesis, and disease outcome.

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ITCH E3 Ubiquitin Ligase Interacts with Ebola Virus VP40 To Regulate Budding

Cited by 62 publications

References 73 publications

Ubiquitin Ligase WWP1 Interacts with Ebola Virus VP40 To Regulate Egress

Ubiquitin Ligase WWP1 Interacts with Ebola Virus VP40 To Regulate Egress

Regulation of Ebola virus VP40 matrix protein by SUMO

Host Factors Involved in Ebola Virus Replication

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