scholarly journals Cells Expressing the RING Finger Z Protein Are Resistant to Arenavirus Infection

2004 ◽  
Vol 78 (6) ◽  
pp. 2979-2983 ◽  
Author(s):  
Tatjana I. Cornu ◽  
Heinz Feldmann ◽  
Juan Carlos de la Torre
Keyword(s):  
Recombinant Adenovirus ◽  
Ring Finger ◽  
Hemorrhagic Fever ◽  
Lassa Fever ◽  
Lymphocytic Choriomeningitis ◽  
Surface Glycoprotein ◽  
Protein Z ◽  
Virus Transcription ◽  
Lassa Fever Virus ◽  
Z Protein

ABSTRACT Arenaviruses include Lassa fever virus (LFV) and the South American hemorrhagic fever viruses. These viruses cause severe human disease, and they pose a threat as agents of bioterrorism. Arenaviruses are enveloped viruses with a bisegmented negative-strand RNA genome whose proteomic capability is limited to four polypeptides: nucleoprotein (NP); surface glycoprotein (GP), which is proteolytically processed into GP1 and GP2; polymerase (L); and a small (11-kDa) RING finger protein (Z). Our investigators have previously shown that Z has a strong inhibitory activity on RNA synthesis mediated by the polymerase of the prototypic arenavirus, lymphocytic choriomeningitis virus (LCMV). In this report we show that cells transduced with a replication-deficient recombinant adenovirus expressing Z (rAd-Z) are resistant to LCMV and LFV infection. Virus cell entry mediated by LCMV or LFV GP was not affected in rAd-Z-transduced cells, but both virus transcription and replication were strongly and specifically inhibited, which resulted in a dramatic reduction in production of infectious virus. These findings open new avenues for developing antiviral strategies to combat the highly pathogenic human arenaviruses, including LFV.

scholarly journals Arenavirus Z-Glycoprotein Association Requires Z Myristoylation but Not Functional RING or Late Domains

2007 ◽  
Vol 81 (17) ◽  
pp. 9451-9460 ◽  
Author(s):  
Althea A. Capul ◽  
Mar Perez ◽  
Emily Burke ◽  
Stefan Kunz ◽  
Michael J. Buchmeier ◽  
...  
Keyword(s):  
Function Analysis ◽  
Ring Finger ◽  
Hemorrhagic Fever ◽  
Lassa Fever ◽  
Lymphocytic Choriomeningitis ◽  
Viral Envelope ◽  
Surface Glycoprotein ◽  
Glycoprotein Precursor ◽  
Biochemical Interaction ◽  
Lassa Fever Virus

ABSTRACT Generation of infectious arenavirus-like particles requires the virus RING finger Z protein and surface glycoprotein precursor (GPC) and the correct processing of GPC into GP1, GP2, and a stable signal peptide (SSP). Z is the driving force of arenavirus budding, whereas the GP complex (GPc), consisting of hetero-oligomers of SSP, GP1, and GP2, forms the viral envelope spikes that mediate receptor recognition and cell entry. Based on the roles played by Z and GP in the arenavirus life cycle, we hypothesized that Z and the GPc should interact in a manner required for virion formation. Here, using confocal microscopy and coimmunoprecipitation assays, we provide evidence for subcellular colocalization and biochemical interaction, respectively, of Z and the GPc. Our results from mutation-function analysis reveal that Z myristoylation, but not the Z late (L) or RING domain, is required for Z-GPc interaction. Moreover, Z interacted directly with SSP in the absence of other components of the GPc. We obtained similar results with Z and GPC from the prototypical arenavirus lymphocytic choriomeningitis virus and the hemorrhagic fever arenavirus Lassa fever virus.

scholarly journals Myristoylation of the RING Finger Z Protein Is Essential for Arenavirus Budding

2004 ◽  
Vol 78 (20) ◽  
pp. 11443-11448 ◽  
Author(s):  
Mar Perez ◽  
Dori L. Greenwald ◽  
Juan Carlos de La Torre
Keyword(s):  
Ring Finger ◽  
Virus Production ◽  
Lassa Fever ◽  
Lymphocytic Choriomeningitis ◽  
Small Ring ◽  
Acceptor Site ◽  
Metabolic Labeling ◽  
Lassa Fever Virus ◽  
Tyrosine Protein Kinase ◽  
Z Protein

ABSTRACT The arenavirus small RING finger Z protein is the main driving force of arenavirus budding. The primary structure of Z is devoid of hydrophobic transmembrane domains, but both lymphocytic choriomeningitis virus (LCMV) and Lassa fever virus Z proteins accumulate near the inner surface of the plasma membrane and are strongly membrane associated. All known arenavirus Z proteins contain a glycine (G) at position 2, which is a potential acceptor site for a myristoyl moiety. Metabolic labeling showed incorporation of [3H]myristic acid by wild-type Z protein but not by the G2A mutant. The mutation G2A eliminated Z-mediated budding. Likewise, treatment with the myristoylation inhibitor 2-hydroxymyristic acid inhibited Z-mediated budding, eliminated formation of virus-like particles, and caused a dramatic reduction in virus production in LCMV-infected cells. Budding activity was restored in G2A mutant Z proteins by the addition of the myristoylation domain of the tyrosine protein kinase Src to their N termini. These findings indicate N-terminal myristoylation of Z plays a key role in arenavirus budding.

scholarly journals RING Finger Z Protein of Lymphocytic Choriomeningitis Virus (LCMV) Inhibits Transcription and RNA Replication of an LCMV S-Segment Minigenome

2001 ◽  
Vol 75 (19) ◽  
pp. 9415-9426 ◽  
Author(s):  
Tatjana I. Cornu ◽  
Juan Carlos de la Torre
Keyword(s):  
Ring Finger ◽  
Rna Replication ◽  
Genetic System ◽  
Lymphocytic Choriomeningitis ◽  
Lymphocytic Choriomeningitis Virus ◽  
Surface Glycoprotein ◽  
Reverse Genetic System ◽  
Protein Z ◽  
Susceptibility To Infection ◽  
Z Protein

ABSTRACT Arenaviruses have a bisegmented negative-strand RNA genome whose proteomic capability is limited to only four polypeptides, namely, nucleoprotein (NP), surface glycoprotein (GP) that is proteolytically processed into GP1+GP2, polymerase (L), and a small (11-kDa) RING finger protein (Z). The role of Z during the Lymphocytic choriomeningitis virus (LCMV) life cycle is poorly understood. We investigated the function of Z in virus transcription and replication by using a reverse genetic system for the prototypic arenavirus LCMV. This system involves an LCMV minigenome and the minimal viral trans-acting factors (NP and L), expressed from separated cotransfected plasmids. Cotransfection of the Z cDNA strongly inhibited LCMV minigenome expression. The effect required synthesis of Z protein; its magnitude was dose dependent and occurred with levels of Z protein substantially lower than those observed in LCMV-infected cells. Coexpression of Z did not prevent the encapsidation of plasmid supplied minigenome, but it affected both transcription and RNA replication similarly. Mutations in Z that unfolded its RING finger domain eliminated its inhibitory activity, but RING proteins not related to Z did not affect LCMV minigenome expression. Consistent with the minigenome results, cells transiently expressing Z exhibited decreased susceptibility to infection with LCMV.

scholarly journals Characterization of the Arenavirus RING Finger Z Protein Regions Required for Z-Mediated Inhibition of Viral RNA Synthesis

2002 ◽  
Vol 76 (13) ◽  
pp. 6678-6688 ◽  
Author(s):  
Tatjana I. Cornu ◽  
Juan Carlos de la Torre
Keyword(s):  
Inhibitory Activity ◽  
Rna Synthesis ◽  
Structural Integrity ◽  
Ring Finger ◽  
Surface Glycoprotein ◽  
Reverse Genetic System ◽  
Ring Finger Protein ◽  
Protein Z ◽  
Finger Protein ◽  
Z Protein

ABSTRACT The prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) is an enveloped virus with a bisegmented negative-strand RNA genome whose proteomic capability is limited to four polypeptides, namely, nucleoprotein; surface glycoprotein (GP), which is proteolytically processed into GP1 and GP2; polymerase (L); and a small (11-kDa) RING finger protein (Z). Using a reverse genetic system based on the ARM strain of LCMV, we have previously shown that Z has a strong inhibitory activity on LCMV minigenome transcription and RNA replication (T. I. Cornu and J. C. de la Torre, J. Virol. 75:9415-9426, 2001). In the present study, we have identified regions and specific amino acid residues within Z which contribute to its inhibitory activity on RNA synthesis mediated by the LCMV polymerase. Z proteins from different LCMV strains had similar inhibitory activities on the expression of the LCMV ARM minigenome, whereas the Z protein of the genetically more distantly related Tacaribe virus had an approximately 10-fold lower inhibitory activity on ARM minigenome expression. Results from the use of chimera proteins between Z and Xenopus Neuralized, a nonviral RING finger protein, indicated that the structural integrity of the Z RING domain (RD) was required but not sufficient for the inhibitory activity of Z. Serial deletion mutants of the N and C termini of Z showed that the N terminus (residues 1 through 16) and C terminus (residues 79 through 90) do not contribute to the Z inhibitory activity. A highly conserved tryptophan (W) residue located at position 36 in ARM-Z, next to the second conserved cysteine (C) residue of the Z RD, also contributed to the Z inhibitory activity.

scholarly journals Host-Driven Phosphorylation Appears to Regulate the Budding Activity of the Lassa Virus Matrix Protein

Pathogens ◽  
2018 ◽  
Vol 7 (4) ◽  
pp. 97 ◽  
Author(s):  
Christopher Ziegler ◽  
Philip Eisenhauer ◽  
Inessa Manuelyan ◽  
Marion Weir ◽  
Emily Bruce ◽  
...  
Keyword(s):  
Protein Function ◽  
Matrix Protein ◽  
Rna Virus ◽  
Hemorrhagic Fever ◽  
Polymerase Activity ◽  
Lassa Fever ◽  
Lassa Virus ◽  
Protein Z ◽  
Western Africa ◽  
Z Protein

Lassa mammarenavirus (LASV) is an enveloped RNA virus that can cause Lassa fever, an acute hemorrhagic fever syndrome associated with significant morbidity and high rates of fatality in endemic regions of western Africa. The arenavirus matrix protein Z has several functions during the virus life cycle, including coordinating viral assembly, driving the release of new virus particles, regulating viral polymerase activity, and antagonizing the host antiviral response. There is limited knowledge regarding how the various functions of Z are regulated. To investigate possible means of regulation, mass spectrometry was used to identify potential sites of phosphorylation in the LASV Z protein. This analysis revealed that two serines (S18, S98) and one tyrosine (Y97) are phosphorylated in the flexible N- and C-terminal regions of the protein. Notably, two of these sites, Y97 and S98, are located in (Y97) or directly adjacent to (S98) the PPXY late domain, an important motif for virus release. Studies with non-phosphorylatable and phosphomimetic Z proteins revealed that these sites are important regulators of the release of LASV particles and that host-driven, reversible phosphorylation may play an important role in the regulation of LASV Z protein function.

scholarly journals Analysis of Assembly and Budding of Lujo Virus

2015 ◽  
Vol 90 (6) ◽  
pp. 3257-3261 ◽  
Author(s):  
Shuzo Urata ◽  
Jacqueline Weyer ◽  
Nadia Storm ◽  
Yukiko Miyazaki ◽  
Petrus Jansen van Vuren ◽  
...  
Keyword(s):  
Cell Surface ◽  
Matrix Protein ◽  
Hemorrhagic Fever ◽  
Surface Glycoprotein ◽  
Protein Z ◽  
Glycoprotein Precursor ◽  
Cell Surface Glycoprotein ◽  
Particle Release ◽  
Viral Release ◽  
Virus Like Particle

The recently identified arenavirus Lujo virus (LUJV) causes fatal hemorrhagic fever in humans. We analyzed its mechanism of viral release driven by matrix protein Z and the cell surface glycoprotein precursor GPC. The L domains in Z are required for efficient virus-like particle release, but Tsg101, ALIX/AIP1, and Vps4A/B are unnecessary for budding. LUJV GPC is cleaved by site 1 protease (S1P) at the RKLM motif, and treatment with the S1P inhibitor PF-429242 reduced LUJV production.

scholarly journals Host Protein BAG3 is a Negative Regulator of Lassa VLP Egress

Diseases ◽  
2018 ◽  
Vol 6 (3) ◽  
pp. 64 ◽  
Author(s):  
Ziying Han ◽  
Michael Schwoerer ◽  
Philip Hicks ◽  
Jingjing Liang ◽  
Gordon Ruthel ◽  
...  
Keyword(s):  
Defense Mechanism ◽  
Hemorrhagic Fever ◽  
Lassa Fever ◽  
Cellular Protein ◽  
Negative Regulator ◽  
Host Protein ◽  
Ww Domain ◽  
Host Defense Mechanism ◽  
Lassa Fever Virus ◽  
Novel Host

Lassa fever virus (LFV) belongs to the Arenaviridae family and can cause acute hemorrhagic fever in humans. The LFV Z protein plays a central role in virion assembly and egress, such that independent expression of LFV Z leads to the production of virus-like particles (VLPs) that mimic egress of infectious virus. LFV Z contains both PTAP and PPPY L-domain motifs that are known to recruit host proteins that are important for mediating efficient virus egress and spread. The viral PPPY motif is known to interact with specific host WW-domain bearing proteins. Here we identified host WW-domain bearing protein BCL2 Associated Athanogene 3 (BAG3) as a LFV Z PPPY interactor using our proline-rich reading array of WW-domain containing mammalian proteins. BAG3 is a stress-induced molecular co-chaperone that functions to regulate cellular protein homeostasis and cell survival via Chaperone-Assisted Selective Autophagy (CASA). Similar to our previously published findings for the VP40 proteins of Ebola and Marburg viruses, our results using VLP budding assays, BAG3 knockout cells, and confocal microscopy indicate that BAG3 is a WW-domain interactor that negatively regulates egress of LFV Z VLPs, rather than promoting VLP release. Our results suggest that CASA and specifically BAG3 may represent a novel host defense mechanism, whereby BAG3 may dampen egress of several hemorrhagic fever viruses by interacting and interfering with the budding function of viral PPxY-containing matrix proteins.

scholarly journals Old World and Clade C New World Arenaviruses Mimic the Molecular Mechanism of Receptor Recognition Used by α-Dystroglycan's Host-Derived Ligands

2007 ◽  
Vol 81 (11) ◽  
pp. 5685-5695 ◽  
Author(s):  
Jillian M. Rojek ◽  
Christina F. Spiropoulou ◽  
Kevin P. Campbell ◽  
Stefan Kunz
Keyword(s):  
New World ◽  
Critical Role ◽  
Lassa Fever ◽  
Lymphocytic Choriomeningitis ◽  
Old World ◽  
Ecm Proteins ◽  
Receptor Recognition ◽  
Lassa Fever Virus ◽  
Clade C

ABSTRACT α-Dystroglycan (DG) is an important cellular receptor for extracellular matrix (ECM) proteins and also serves as the receptor for Old World arenaviruses Lassa fever virus (LFV) and lymphocytic choriomeningitis virus (LCMV) and clade C New World arenaviruses. In the host cell, α-DG is subject to a remarkably complex pattern of O glycosylation that is crucial for its interactions with ECM proteins. Two of these unusual sugar modifications, protein O mannosylation and glycan modifications involving the putative glycosyltransferase LARGE, have recently been implicated in arenavirus binding. Considering the complexity of α-DG O glycosylation, our present study was aimed at the identification of the specific O-linked glycans on α-DG that are recognized by arenaviruses. As previously shown for LCMV, we found that protein O mannosylation of α-DG is crucial for the binding of arenaviruses of distinct phylogenetic origins, including LFV, Mobala virus, and clade C New World arenaviruses. In contrast to the highly conserved requirement for O mannosylation, more generic O glycans present on α-DG are dispensable for arenavirus binding. Despite the critical role of O-mannosyl glycans for arenavirus binding under normal conditions, the overexpression of LARGE in cells deficient in O mannosylation resulted in highly glycosylated α-DG that was functional as a receptor for arenaviruses. Thus, modifications by LARGE but not O-mannosyl glycans themselves are most likely the crucial structures recognized by arenaviruses. Together, the data demonstrate that arenaviruses recognize the same highly conserved O-glycan structures on α-DG involved in ECM protein binding, indicating a strikingly similar mechanism of receptor recognition by pathogen- and host-derived ligands.

scholarly journals Common Antiviral Cytotoxic T-Lymphocyte Epitope for Diverse Arenaviruses

2001 ◽  
Vol 75 (14) ◽  
pp. 6273-6278 ◽  
Author(s):  
Michael B. A. Oldstone ◽  
Hanna Lewicki ◽  
Dirk Homann ◽  
Christophe Nguyen ◽  
Sylvianne Julien ◽  
...  
Keyword(s):  
Amino Acids ◽  
T Lymphocyte ◽  
New World ◽  
Cytotoxic T Lymphocyte ◽  
Lassa Fever ◽  
Lymphocytic Choriomeningitis ◽  
Target Cells ◽  
Old World ◽  
Ctl Epitope ◽  
Lassa Fever Virus

ABSTRACT Members of the Arenaviridae family have been isolated from mammalian hosts in disparate geographic locations, leading to their grouping as Old World types (i.e., lymphocytic choriomeningitis virus [LCMV], Lassa fever virus [LFV], Mopeia virus, and Mobala virus) and New World types (i.e., Junin, Machupo, Tacaribe, and Sabia viruses) (C. J. Peters, M. J. Buchmeier, P. E. Rollin, and T. G. Ksiazek, p. 1521–1551, in B. N. Fields, D. M. Knipe, and P. M. Howley [ed.], Fields virology, 3rd ed., 1996; P. J. Southern, p. 1505–1519, in B. N. Fields, D. M. Knipe, and P. M. Howley [ed.], Fields virology, 3rd ed., 1996). Several types in both groups—LFV, Junin, Machupo, and Sabia viruses—cause severe and often lethal human diseases. By sequence comparison, we noted that eight Old World and New World arenaviruses share several amino acids with the nucleoprotein (NP) that consists of amino acids (aa) 118 to 126 (NP 118–126) (RPQASGVYM) of LCMV that comprise the immunodominant cytotoxic T-lymphocyte (CTL) epitope forH-2 d mice (32). This Ld-restricted epitope constituted >97% of the total bulk CTLs produced in the specific antiviral or clonal responses ofH-2 d BALB mice. NP 118–126 of the Old World arenaviruses LFV, Mopeia virus, and LCMV and the New World arenavirus Sabia virus bound at high affinity to Ld. The primary H-2 d CTL anti-LCMV response as well as that of a CTL clone responsive to LCMV NP 118–126 recognized target cells coated with NP 118–126 peptides derived from LCMV, LFV, and Mopeia virus but not Sabia virus, indicating that a common functional NP epitope exists among Old World arenaviruses. Use of site-specific amino acid exchanges in the NP CTL epitope among these arenaviruses identified amino acids involved in major histocompatibility complex binding and CTL recognition.

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