Membrane-associated RING-CH (MARCH) 1 and 2 are MARCH family members that inhibit HIV-1 infection

Zhang, Yanzhao; Tada, Takuya; Ozono, Seiya; Yao, Weitong; Tanaka, Michiko; Shoji, Yasuhiro; Kishigami, Satoshi; Fujita, Hideaki; Tokunaga, Kenzo

doi:10.1074/jbc.ac118.005907

Cited by 51 publications

(94 citation statements)

References 32 publications

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“…On the other hand, the mutant MARCH8 had a completely abrogated ability to downregulate cell-surface HIV-1 Env, whereas WT expression led to the downregulation of HIV-1 Env from the cell surface and its intracellular retention (Fig. 2H), as we previously observed 12 . The results were consistent with those of the inhibitory activity of MARCH8 against the virion incorporation of HIV-1 Env (Fig.…”

Section: Resultssupporting

confidence: 81%

“…In terms of the latter mechanism, although this could be attributed to the AP-dependent trafficking that requires the YxxΦ motif, which binds to AP μ-subunits, we were unable to prove the interaction between these subunits and MARCH8 (data not shown). We have recently reported that MARCH1 and MARCH2 are also antiviral MARCH family members that inhibit HIV-1 infection, although their antiviral activity is less robust, which is probably due to the lower protein expression and/or stability than that of MARCH8 12 . Because MARCH1 and MARCH2 also harbor the Yxxϕ motif in their C-terminal CT domains, it would be intriguing to verify the importance of the motif in these proteins.…”

Section: Resultsmentioning

confidence: 99%

“…Protein expression of constructs was confirmed by Western blot analyses as described elsewhere 10,12 . Briefly, cells transfected as described above were lysed in 500 μl of lysis buffer containing 1.25% n-octyl-β-D-glucoside, and Complete protease inhibitor mixture (Roche Applied Science).…”

Section: Methodsmentioning

confidence: 99%

“…A β-lactamase (BlaM)-fused viral protein R (Vpr)-based entry assay was performed as described elsewhere 10,12 . Briefly, HIV-1 particles containing a fusion protein of Vpr and BlaM-Vpr were produced by cotransfection of 293T cells with pNL4-3, pMM310 14 encoding BlaM-Vpr, and either pC-MARCH8, pC-MARCH8- 222 AxxL 225 , pC-MARCH8- 232 AxxV 235 , or the control vector.…”

Section: Methodsmentioning

confidence: 99%

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MARCH8 inhibits viral infection by two different mechanisms

Zhang

Tada

Ozono

et al. 2020

Preprint

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Membrane-associated RING-CH 8 (MARCH8) inhibits infection with both HIV-1 and vesicular stomatitis virus G-glycoprotein (VSV-G)-pseudotyped viruses by reducing virion incorporation of envelope glycoproteins. The molecular mechanisms by which MARCH8 targets envelope glycoproteins remain unknown. Here, we show two different mechanisms by which MARCH8 inhibits viral infection. Viruses pseudotyped with the VSV-G mutant, in which cytoplasmic lysine residues were mutated, were insensitive to the inhibitory effect of MARCH8, whereas those with a similar lysine mutant of HIV-1 Env remained sensitive to it. Indeed, the wild-type VSV-G, but not its lysine mutant, was ubiquitinated by MARCH8. Furthermore, the MARCH8 mutant, which had a disrupted cytoplasmic tyrosine motif that is critical for intracellular protein sorting, did not inhibit HIV-1 Env-mediated infection, while it still impaired infection by VSV-G-pseudotyped viruses. Overall, we conclude that MARCH8 reduces viral infectivity by downregulating envelope glycoproteins through two different mechanisms mediated by a ubiquitination-dependent or tyrosine motif-dependent pathway.

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Section: Resultssupporting

confidence: 81%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

MARCH8 inhibits viral infection by two different mechanisms

Zhang

Tada

Ozono

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…MARCH8 is the first identified cellular MIR (c-MIR) from the human genome ( 12 , 13 ), and MARCH proteins have been found to downregulate a number of cellular proteins from the cell surface ( 14 ). Recently, MARCH8 was found to block HIV-1 Env incorporation and inhibit viral replication by downregulating Env from the cell surface ( 15 , 16 ). We now report that MARCH8 has a very broad antiviral activity that inhibits EBOV GP, HIV-1 Env, and H5N1 HA maturation.…”

Section: Introductionmentioning

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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Human MARCH1, 2, and 8 block Ebola virus envelope glycoprotein cleavage via targeting furin P domain

Yu,

Bai,

Tan

et al. 2024

Journal of Medical Virology

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Membrane‐associated RING‐CH (MARCH) family proteins were recently reported to inhibit viral replication through multiple modes. Previous work showed that human MARCH8 blocked Ebola virus (EBOV) glycoprotein (GP) maturation. Our study here demonstrates that human MARCH1 and MARCH2 share a similar pattern to MARCH8 in restricting EBOV GP‐pseudotyped viral infection. Human MARCH1 and MARCH2 retain EBOV GP at the trans‐Golgi network, reduce its cell surface display, and impair EBOV GP‐pseudotyped virions infectivity. Furthermore, we uncover that the host proprotein convertase furin could interact with human MARCH1/2 and EBOV GP intracellularly. Importantly, the furin P domain is verified to be recognized by MARCH1/2/8, which is critical for their blocking activities. Besides, bovine MARCH2 and murine MARCH1 also impair EBOV GP proteolytic processing. Altogether, our findings confirm that MARCH1/2 proteins of different mammalian origins showed a relatively conserved feature in blocking EBOV GP cleavage, which could provide clues for subsequent MARCHs antiviral studies and may facilitate the development of novel strategies to antagonize enveloped virus infection.

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Membrane-associated RING-CH (MARCH) 1 and 2 are MARCH family members that inhibit HIV-1 infection

Cited by 51 publications

References 32 publications

MARCH8 inhibits viral infection by two different mechanisms

MARCH8 inhibits viral infection by two different mechanisms

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

Human MARCH1, 2, and 8 block Ebola virus envelope glycoprotein cleavage via targeting furin P domain

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