MARCH8 inhibits viral infection by two different mechanisms

Zhang, Yanzhao; Tada, Takuya; Ozono, Seiya; Kishigami, Satoshi; Fujita, Hideaki; Tokunaga, Kenzo

doi:10.1101/2020.04.09.035204

Cited by 8 publications

(19 citation statements)

References 15 publications

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“…In all cases, however, the ubiquitin ligase activity of MARCH8 was required for restriction, as RING domain mutations to a large extent abrogated the inhibition. While this manuscript was in preparation, another group reported that, as we observed here, MARCH8-mediated restriction of VSV-G was CT-dependent but antagonism of HIV-1 Env was CT-independent, and they demonstrated MARCH8-mediated ubiquitination of VSV-G(55). These results support a model whereby the RING domain E3 ubiquitin ligase activity of MARCH8 either directly targets the CT of the viral glycoproteinas has been shown for MARCH protein-mediated downregulation of a variety of cellular proteins (18)or exerts its effect indirectly.…”

supporting

confidence: 66%

“…Our results show a broad antiviral activity of MARCH8 against the glycoproteins from three human viral pathogens – HIV-1, EboV, and SARS-CoV-2 – and a primarily animal pathogen, VSV. MARCH8 targeting of HIV-1 Env, VSV-G, and, very recently, EboV-GP, has been reported (6, 8, 11, 55, 59). HIV-1 replicates predominantly in CD4 + T cells and also infects MDM (60).…”

Section: Discussionmentioning

confidence: 99%

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Mechanism of Viral Glycoprotein Targeting by Membrane-associated-RING-CH Proteins

Lun

Waheed

Majadly

et al. 2021

Preprint

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An emerging class of cellular inhibitory proteins has been identified that targets viral glycoproteins. These include the membrane-associated RING-CH (MARCH) family of E3 ubiquitin ligases that, among other functions, downregulate cell-surface proteins involved in adaptive immunity. The RING-CH domain of MARCH proteins is thought to function by catalyzing the ubiquitination of the cytoplasmic tails (CTs) of target proteins, leading to their degradation. MARCH proteins have recently been reported to target retroviral envelope glycoproteins (Env) and vesicular stomatitis virus G glycoprotein (VSV-G). However, the mechanism of antiviral activity remains poorly defined. Here we show that MARCH8 antagonizes the full-length forms of HIV-1 Env, VSV-G, Ebola virus glycoprotein (EboV-GP), and the spike (S) protein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) thereby impairing the infectivity of virions pseudotyped with these viral glycoproteins. This MARCH8-mediated targeting of viral glycoproteins requires the E3 ubiquitin ligase activity of the RING-CH domain. We observe that MARCH8 protein antagonism of VSV-G is CT dependent. In contrast, MARCH8-mediated targeting of HIV-1 Env, EboV-GP and SARS-CoV-2 S protein by MARCH8 does not require the CT, suggesting a novel mechanism of MARCH-mediated antagonism of these viral glycoproteins. Confocal microscopy data demonstrate that MARCH8 traps the viral glycoproteins in an intracellular compartment. We observe that the endogenous expression of MARCH8 in several relevant human cell types is rapidly inducible by type I interferon. These results help to inform the mechanism by which MARCH proteins exert their antiviral activity and provide insights into the role of cellular inhibitory factors in antagonizing the biogenesis, trafficking, and virion incorporation of viral glycoproteins.ImportanceViral envelope glycoproteins are an important structural component on the surface of enveloped viruses that direct virus binding and entry and also serve as targets for the host adaptive immune response. In this study, we investigate the mechanism of action of the MARCH family of cellular proteins that disrupt the trafficking and virion incorporation of viral glycoproteins across several virus families. This research provides novel insights into how host cell factors antagonize viral replication, perhaps opening new avenues for therapeutic intervention in the replication of a diverse group of highly pathogenic enveloped viruses.

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supporting

confidence: 66%

Section: Discussionmentioning

confidence: 99%

Mechanism of Viral Glycoprotein Targeting by Membrane-associated-RING-CH Proteins

Lun

Waheed

Majadly

et al. 2021

Preprint

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“…We speculate that MARCH8 might ubiquitinate and degrade a critical cellular transport facilitator, which blocks GP from access to the glycosylation and furin cleavage machinery. However, the degradation of VSV-G by MARCH8 follows a similar mechanism by which MARCH proteins downregulate immune receptors from the cell surface ( 29 ). MARCH proteins usually ubiquitinate a lysine residue in the cytoplasmic tail of these immune receptors, which redirects them from the endosomes to the late endosomes and the lysosomes for degradation when they are endocytosed from the cell surface ( 14 ).…”

Section: Discussionmentioning

confidence: 99%

MARCH8 Inhibits Ebola Virus Glycoprotein, Human Immunodeficiency Virus Type 1 Envelope Glycoprotein, and Avian Influenza Virus H5N1 Hemagglutinin Maturation

Zhang

et al. 2020

mBio

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Membrane-associated RING-CH-type 8 (MARCH8) strongly blocks human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) incorporation into virions by downregulating its cell surface expression, but the mechanism is still unclear. We now report that MARCH8 also blocks the Ebola virus (EBOV) glycoprotein (GP) incorporation via surface downregulation. To understand how these viral fusion proteins are downregulated, we investigated the effects of MARCH8 on EBOV GP maturation and externalization via the conventional secretion pathway. MARCH8 interacted with EBOV GP and furin when detected by immunoprecipitation and retained the GP/furin complex in the Golgi when their location was tracked by a bimolecular fluorescence complementation (BiFC) assay. MARCH8 did not reduce the GP expression or affect the GP modification by high-mannose N-glycans in the endoplasmic reticulum (ER), but it inhibited the formation of complex N-glycans on the GP in the Golgi. Additionally, the GP O-glycosylation and furin-mediated proteolytic cleavage were also inhibited. Moreover, we identified a novel furin cleavage site on EBOV GP and found that only those fully glycosylated GPs were processed by furin and incorporated into virions. Furthermore, the GP shedding and secretion were all blocked by MARCH8. MARCH8 also blocked the furin-mediated cleavage of HIV-1 Env (gp160) and the highly pathogenic avian influenza virus H5N1 hemagglutinin (HA). We conclude that MARCH8 has a very broad antiviral activity by prohibiting different viral fusion proteins from glycosylation and proteolytic cleavage in the Golgi, which inhibits their transport from the Golgi to the plasma membrane and incorporation into virions. IMPORTANCE Enveloped viruses express three classes of fusion proteins that are required for their entry into host cells via mediating virus and cell membrane fusion. Class I fusion proteins are produced from influenza viruses, retroviruses, Ebola viruses, and coronaviruses. They are first synthesized as a type I transmembrane polypeptide precursor that is subsequently glycosylated and oligomerized. Most of these precursors are cleaved en route to the plasma membrane by a cellular protease furin in the late secretory pathway, generating the trimeric N-terminal receptor-binding and C-terminal fusion subunits. Here, we show that a cellular protein, MARCH8, specifically inhibits the furin-mediated cleavage of EBOV GP, HIV-1 Env, and H5N1 HA. Further analyses uncovered that MARCH8 blocked the EBOV GP glycosylation in the Golgi and inhibited its transport from the Golgi to the plasma membrane. Thus, MARCH8 has a very broad antiviral activity by specifically inactivating different viral fusion proteins.

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“…Besides, the MARCH8 mutant, with a disrupted cytoplasmic tyrosine motif essential for intracellular protein sorting, did not inhibit HIV-1 Env-mediated infection while still affecting VSV-G-pseudotyped virus infection. The underlying mechanism is that MARCH8 decreases viral infectivity by downregulating envelope glycoproteins through two separate mechanisms mediated by a ubiquitinationdependent or tyrosine motif-dependent pathway [21]. Further investigations are required to clarify the more comprehensive host defense mechanisms for MRACH8.…”

Section: March8mentioning

confidence: 99%

When MARCH family proteins meet viral infections

Zheng

Tang

2021

Virol J

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Membrane-associated RING-CH (MARCH) ubiquitin ligases belong to a RING finger domain E3 ligases family. Recent studies have demonstrated that MARCH proteins play critical roles during various viral infections. MARCH proteins can directly antagonize different steps of the viral life cycle and promote individual viral infection. This mini-review will focus on the latest advances of MARCH family proteins' emerging roles during viral infections.

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MARCH8 inhibits viral infection by two different mechanisms

Cited by 8 publications

References 15 publications

Mechanism of Viral Glycoprotein Targeting by Membrane-associated-RING-CH Proteins

Mechanism of Viral Glycoprotein Targeting by Membrane-associated-RING-CH Proteins

MARCH8 Inhibits Ebola Virus Glycoprotein, Human Immunodeficiency Virus Type 1 Envelope Glycoprotein, and Avian Influenza Virus H5N1 Hemagglutinin Maturation

When MARCH family proteins meet viral infections

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