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1.
Cell Biol Int ; 2024 Jul 15.
Article in English | MEDLINE | ID: mdl-39010645

ABSTRACT

Migrasome is a newly discovered organelle composed of small vesicular structures enclosed in membrane structures. Since its discovery in 2014, migrasome has attracted increasing attention in cell biology due to its critical role in multiple disease processes. Its pivotal role in various disease processes, including cell migration, intercellular communication, removal of damaged mitochondria, embryogenesis localization, immune cell chemotaxis, and virus transmission, underscores its significance in biological systems. With research on migrasome steadily increasing, it becomes a unique resource for undergraduate cell biology education. For deeper understanding of migrasome, we applied a bibliometric approach. Here we conducted a comprehensive analysis of migrasome research by retrieving relevant literature from databases such as Web of Science, Scopus, and PubMed using the keywords "migrasome" or "migrasomes." Employing CiteSpace software and Prism, we analyzed annual publication trends, identified core authors and institutions, assessed national contributions, examined keywords, and scrutinized highly cited literature related to migrasome research. This study presents a comprehensive overview of migrasome research, elucidating its literature characteristics, key contributors, research hotspots, and emerging trends. By shedding light on the current status and future trajectories of migrasome research, we aim to provide valuable insights for teachers in cell biology education. We propose for the integration of migrasome research into undergraduate curricula to enhance the understanding of cell biology among premedical, medical, and biomedical students, thereby fostering a deeper appreciation for the intricate mechanisms governing cellular behavior and disease processes.

2.
J Med Virol ; 95(1): e28299, 2023 01.
Article in English | MEDLINE | ID: mdl-36366981

ABSTRACT

Monkeypox virus (MPXV) is a member of Orthopoxvirus in the Poxviridae family, causing a Public Health Emergency of International Concern. The number of cases and geographic range has increased significantly in 2022. Identification of MPXV-specific therapeutic targets is urgent. G-quadruplex (GQ) secondary structures attract great attention as potential targets for antiviral strategy. Whether GQs are present in the MPXV genome remains inconclusive. In this study, we aim to characterize the GQs encoded by MPXV. Through a series of biophysical experiments, we characterized the formation potential of MPXV-encoded GQs and evaluated the binding and stabilization abilities of GQ ligands including BRACO-19, pyridostatin, and TMPyP4 to GQs encoded by MPXV. Moreover, GQ ligands suppressed the gene transcription of MPXV sequences containing GQ. BRACO-19 and TMPyP4 were able to inhibit vaccinia virus replication. We demonstrated the existence of MPXV GQ and reinforced the idea that GQs could be novel antiviral targets. Targeting these GQ sequences with GQ-binding molecules may represent a new approach for MPXV therapy.


Subject(s)
G-Quadruplexes , Mpox (monkeypox) , Humans , Monkeypox virus/genetics , Antiviral Agents/pharmacology , Ligands
3.
J Med Virol ; 95(2): e28513, 2023 02.
Article in English | MEDLINE | ID: mdl-36661039

ABSTRACT

Mpox is caused by the mpox virus, which belongs to the Orthopoxvirus genus and Poxviridae family. Animal hosts, such as African rodents, mice, prairie dogs, and non-human primates, play important roles in the development and transmission of outbreaks. Laboratory animal infection experiments have demonstrated that some animals are susceptible to mpox virus. This review summarizes the current progress on the animal hosts for mpox virus. The surveillance of mpox virus in animal hosts will provide important insights into virus tracing, analysis of mutation evolutionary patterns, transmission mechanisms, and development of control measures.


Subject(s)
Monkeypox virus , Mpox (monkeypox) , Animals , Mice , Host Specificity , Primates/virology , Sciuridae/virology , Mpox (monkeypox)/veterinary
4.
Adv Exp Med Biol ; 1407: 313-327, 2023.
Article in English | MEDLINE | ID: mdl-36920705

ABSTRACT

Members of Flaviviridae are enveloped single positive-stranded RNA viruses including hepacivirus, pestivirus, pegivirus, and mosquito-transmitted flavivirus, which are important pathogens of infectious diseases and pose serious threats to human health. Pseudotyped virus is an artificially constructed virus-like particle, which could infect host cells similar to a live virus but cannot produce infectious progeny virus. Therefore, pseudotyped virus has the advantages of a wide host range, high transfection efficiency, low biosafety risk, and accurate and objective quantification. It has been widely used in biological characteristics, drug screening, detection methods, and vaccine evaluation of Flaviviridae viruses like hepatitis C virus, Japanese encephalitis virus, dengue virus, and Zika virus.


Subject(s)
Flaviviridae , Flavivirus , Zika Virus Infection , Zika Virus , Animals , Humans , Flaviviridae/genetics , Viral Pseudotyping , Flavivirus/genetics , Hepacivirus/genetics , Zika Virus/genetics
5.
J Med Virol ; 94(1): 342-348, 2022 01.
Article in English | MEDLINE | ID: mdl-34528721

ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of the ongoing coronavirus disease 2019 (COVID-19) pandemic. The S protein is the key viral protein for associating with ACE2, the receptor for SARS-CoV-2. There are many kinds of posttranslational modifications in S protein. However, the detailed mechanism of palmitoylation of SARS-CoV-2 S remains to be elucidated. In our current study, we characterized the palmitoylation of SARS-CoV-2 S. Both the C15 and cytoplasmic tail of SARS-CoV-2 S were palmitoylated. Fatty acid synthase inhibitor C75 and zinc finger DHHC domain-containing palmitoyltransferase (ZDHHC) inhibitor 2-BP reduced the palmitoylation of S. Interestingly, palmitoylation of SARS-CoV-2 S was not required for plasma membrane targeting of S but was critical for S-mediated syncytia formation and SARS-CoV-2 pseudovirus particle entry. Overexpression of ZDHHC2, ZDHHC3, ZDHHC4, ZDHHC5, ZDHHC8, ZDHHC9, ZDHHC11, ZDHHC14, ZDHHC16, ZDHHC19, and ZDHHC20 promoted the palmitoylation of S. Furthermore, those ZDHHCs were identified to associate with SARS-CoV-2 S. Our study not only reveals the mechanism of S palmitoylation but also will shed important light into the role of S palmitoylation in syncytia formation and virus entry.


Subject(s)
Cell Membrane/metabolism , Giant Cells/metabolism , Lipoylation/physiology , Spike Glycoprotein, Coronavirus/metabolism , Virus Internalization , 4-Butyrolactone/analogs & derivatives , 4-Butyrolactone/pharmacology , Acyltransferases/antagonists & inhibitors , COVID-19/pathology , Cell Line , HEK293 Cells , Humans , Protein Processing, Post-Translational/physiology
6.
J Med Virol ; 94(6): 2519-2527, 2022 06.
Article in English | MEDLINE | ID: mdl-35075669

ABSTRACT

Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus affecting human health globally. G-quadruplex secondary structures attract great attention as potential targets for antiviral strategy. In this study, we show that the CHIKV genome possesses several conserved potential G-quadruplex sequences. G-quadruplex ligands BRACO-19 and TMPyP4 could stabilize the CHIKV G-quadruplex and inhibit the transcription of constructs containing CHIKV G-quadruplex sequences. Importantly, BRACO-19 and TMPyP4 suppress CHIKV replication. Our study not only reinforces the presence of viral G-quadruplex sequences but also suggests that targeting G-quadruplex structure could represent a novel strategy to inhibit CHIKV.


Subject(s)
Chikungunya Fever , Chikungunya virus , Animals , Antiviral Agents/pharmacology , Chikungunya virus/genetics , Humans , Ligands , Virus Replication
7.
Biochem Biophys Res Commun ; 538: 72-79, 2021 01 29.
Article in English | MEDLINE | ID: mdl-33276953

ABSTRACT

SARS-CoV-2 papain-like protease is considered as an important potential target for anti-SARS-CoV-2 drug discovery due to its crucial roles in viral spread and innate immunity. Here, we have utilized an in silico molecular docking approach to identify the possible inhibitors of the SARS-CoV-2 papain-like protease, by screening 21 antiviral, antifungal and anticancer compounds. Among them, Neobavaisoflavone has the highest binding energy for SARS-CoV-2 papain-like protease. These molecules could bind near the SARS-CoV-2 papain-like protease crucial catalytic triad, ubiquitination and ISGylation residues: Trp106, Asn109, Cys111, Met208, Lys232, Pro247, Tyr268, Gln269, His272, Asp286 and Thr301. Because blocking the papain-like protease is an important strategy in fighting against viruses, these compounds might be promising candidates for therapeutic intervention against COVID-19.


Subject(s)
Coronavirus Papain-Like Proteases/chemistry , Coronavirus Protease Inhibitors/chemistry , Cysteine Proteinase Inhibitors/chemistry , Drug Discovery/methods , Isoflavones/chemistry , SARS-CoV-2/drug effects , SARS-CoV-2/enzymology , Coronavirus Papain-Like Proteases/antagonists & inhibitors , Coronavirus Protease Inhibitors/pharmacology , Cysteine Proteinase Inhibitors/pharmacology , Humans , Isoflavones/pharmacology , Ligands , Molecular Docking Simulation , Protein Binding
8.
Virus Genes ; 57(6): 475-488, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34510331

ABSTRACT

Throughout the viral life cycle, interplays between cellular host factors and virus determine the infectious capacity of the virus. The pandemic of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) poses a great threat to human life and health. Extensive studies identified a number of host proviral and antiviral factors for SARS-CoV-2. In this review, we summarize the current understanding of the interplay between SARS-CoV-2 and cellular factors during virus entry and replication. Our review will highlight the future direction of study on the infection and pathogenesis of SARS-CoV-2, as well as novel therapeutic strategies and effective antiviral targets for COVID-19.


Subject(s)
COVID-19/metabolism , COVID-19/virology , Host Microbial Interactions/physiology , SARS-CoV-2/physiology , Virus Internalization , Virus Replication , Animals , Humans , SARS-CoV-2/pathogenicity
9.
Nucleic Acids Res ; 47(16): 8693-8707, 2019 09 19.
Article in English | MEDLINE | ID: mdl-31361901

ABSTRACT

Zika virus is a positive single-strand RNA virus whose replication involved RNA unwinding and synthesis. ZIKV NS3 contains a helicase domain, but its enzymatic activity is not fully characterized. Here, we established a dsRNA unwinding assay based on the FRET effect to study the helicase activity of ZIKV NS3, which provided kinetic information in real time. We found that ZIKV NS3 specifically unwound dsRNA/dsDNA with a 3' overhang in the 3' to 5' direction. The RNA unwinding ability of NS3 significantly decreased when the duplex was longer than 18 base pairs. The helicase activity of NS3 depends on ATP hydrolysis and binding to RNA. Mutations in the ATP binding region or the RNA binding region of NS3 impair its helicase activity, thus blocking viral replication in the cell. Furthermore, we showed that ZIKV NS5 interacted with NS3 and stimulated its helicase activity. Disrupting NS3-NS5 interaction resulted in a defect in viral replication, revealing the tight coupling of RNA unwinding and synthesis. We suggest that NS3 helicase activity is stimulated by NS5; thus, viral replication can be carried out efficiently. Our work provides a molecular mechanism of ZIKV NS3 unwinding and novel insights into ZIKV replication.


Subject(s)
Gene Expression Regulation, Viral , RNA, Double-Stranded/chemistry , RNA, Viral/chemistry , Viral Nonstructural Proteins/chemistry , Zika Virus/genetics , Adenosine Triphosphate/chemistry , Adenosine Triphosphate/metabolism , Animals , Binding Sites , Chlorocebus aethiops , Cloning, Molecular , Cricetulus , Epithelial Cells/virology , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Genetic Vectors/chemistry , Genetic Vectors/metabolism , Kinetics , Models, Molecular , Protein Binding , Protein Conformation, alpha-Helical , Protein Conformation, beta-Strand , Protein Interaction Domains and Motifs , Protein Structure, Tertiary , RNA Helicases/chemistry , RNA Helicases/genetics , RNA Helicases/metabolism , RNA, Double-Stranded/genetics , RNA, Double-Stranded/metabolism , RNA, Viral/genetics , RNA, Viral/metabolism , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Serine Endopeptidases/chemistry , Serine Endopeptidases/genetics , Serine Endopeptidases/metabolism , Substrate Specificity , Vero Cells , Viral Nonstructural Proteins/genetics , Viral Nonstructural Proteins/metabolism , Zika Virus/metabolism
10.
Biochem Biophys Res Commun ; 526(1): 165-169, 2020 05 21.
Article in English | MEDLINE | ID: mdl-32201080

ABSTRACT

SARS-CoV-2 causes the recent global COVID-19 public health emergency. ACE2 is the receptor for both SARS-CoV-2 and SARS-CoV. To predict the potential host range of SARS-CoV-2, we analyzed the key residues of ACE2 for recognizing S protein. We found that most of the selected mammals including pets (dog and cat), pangolin and Circetidae mammals remained the most of key residues for association with S protein from SARS-CoV and SARS-CoV-2. The interaction interface between cat/dog/pangolin/Chinese hamster ACE2 and SARS-CoV/SARS-CoV-2 S protein was simulated through homology modeling. We identified that N82 in ACE2 showed a closer contact with SARS-CoV-2 S protein than M82 in human ACE2. Our finding will provide important insights into the host range of SARS-CoV-2 and a new strategy to design an optimized ACE2 for SARS-CoV-2 infection.


Subject(s)
Betacoronavirus/physiology , Peptidyl-Dipeptidase A/metabolism , Spike Glycoprotein, Coronavirus/metabolism , Viral Tropism , Amino Acid Sequence , Angiotensin-Converting Enzyme 2 , Animals , COVID-19 , Coronavirus Infections/metabolism , Coronavirus Infections/transmission , Coronavirus Infections/virology , Humans , Mammals/classification , Mammals/metabolism , Models, Molecular , Pandemics , Peptidyl-Dipeptidase A/chemistry , Pneumonia, Viral/metabolism , Pneumonia, Viral/transmission , Pneumonia, Viral/virology , SARS-CoV-2 , Sequence Alignment , Spike Glycoprotein, Coronavirus/chemistry
11.
J Virol ; 93(3)2019 02 01.
Article in English | MEDLINE | ID: mdl-30404808

ABSTRACT

Chikungunya virus (CHIKV) is transmitted to people by mosquitoes, and CHIKV infection causes fever and joint pain. Fatty acid synthase (FASN) has been identified as a proviral factor for CHIKV. How FASN participates in CHIKV replication remains to be elucidated. In this study, we demonstrated that palmitic acid (PA) can restore the suppression of CHIKV replication by FASN inhibitors. The palmitoylation and plasma membrane localization of CHIKV nsP1 were reduced by FASN inhibitors. Triple mutation of Cys417, Cys418, and Cys419 in nsP1 blocked its palmitoylation and severely disrupted CHIKV replication. Furthermore, two zinc finger DHHC domain-containing palmitoyltransferases (ZDHHCs), ZDHHC2 and ZDHHC19, promoted nsP1 palmitoylation and CHIKV replication. Our results not only identified the key enzymes for the palmitoylation of nsP1 but also provided mechanistic insights into the roles of FASN in CHIKV replication.IMPORTANCE S-palmitoylation is an important form of lipid posttranslational modification, which affects the function of proteins by regulating their transport, stability, and localization. Previous studies have shown that FASN is critical for CHIKV replication; however, the mechanism for this function of FASN remains unknown. The key zinc finger DHHC domain-containing palmitoyltransferases involved in the palmitoylation of nsP1 are not clear. We demonstrated that FASN promoted CHIKV replication through nsP1 palmitoylation. ZDHHC2 and ZDHHC19 were identified as the major enzymes for nsP1 palmitoylation. Since nsP1 proteins are conserved in alphaviruses, our results highlight the mechanisms by which alphavirus nsP1 is palmitoylated.


Subject(s)
Chikungunya Fever/virology , Chikungunya virus/pathogenicity , Fatty Acid Synthase, Type I/metabolism , Lipoylation , Palmitic Acid/metabolism , Viral Nonstructural Proteins/metabolism , Virus Replication , Acyltransferases/genetics , Acyltransferases/metabolism , Chikungunya Fever/genetics , Chikungunya Fever/metabolism , Fatty Acid Synthase, Type I/genetics , HeLa Cells , Humans , Tumor Suppressor Proteins/genetics , Tumor Suppressor Proteins/metabolism , Viral Nonstructural Proteins/genetics
12.
J Virol ; 93(1)2019 01 01.
Article in English | MEDLINE | ID: mdl-30333168

ABSTRACT

Type I interferon (IFN) inhibits viruses by inducing the expression of antiviral proteins. The IFN-induced myxovirus resistance B (MxB) protein has been reported to inhibit a limited number of viruses, including HIV-1 and herpesviruses, but its antiviral coverage remains to be explored further. Here we show that MxB interferes with RNA replication of hepatitis C virus (HCV) and significantly inhibits viral replication in a cyclophilin A (CypA)-dependent manner. Our data further show that MxB interacts with the HCV protein NS5A, thereby impairing NS5A interaction with CypA and NS5A localization to the endoplasmic reticulum, two events essential for HCV RNA replication. Interestingly, we found that MxB significantly inhibits two additional CypA-dependent viruses of the Flaviviridae family, namely, Japanese encephalitis virus and dengue virus, suggesting a potential link between virus dependence on CypA and virus susceptibility to MxB inhibition. Collectively, these data have identified MxB as a key factor behind IFN-mediated suppression of HCV infection, and they suggest that other CypA-dependent viruses may also be subjected to MxB restriction.IMPORTANCE Viruses of the Flaviviridae family cause major illness and death around the world and thus pose a great threat to human health. Here we show that IFN-inducible MxB restricts several members of the Flaviviridae, including HCV, Japanese encephalitis virus, and dengue virus. This finding not only suggests an active role of MxB in combating these major pathogenic human viruses but also significantly expands the antiviral spectrum of MxB. Our study further strengthens the link between virus dependence on CypA and susceptibility to MxB restriction and also suggests that MxB may employ a common mechanism to inhibit different viruses. Elucidating the antiviral functions of MxB advances our understanding of IFN-mediated host antiviral defense and may open new avenues to the development of novel antiviral therapeutics.


Subject(s)
Cyclophilin A/pharmacology , Hepacivirus/physiology , Interferons/pharmacology , Myxovirus Resistance Proteins/metabolism , Viral Nonstructural Proteins/metabolism , Virus Replication/drug effects , Animals , Cell Line , Chlorocebus aethiops , Cyclosporine/pharmacology , Endoplasmic Reticulum/metabolism , Gene Knockdown Techniques , HEK293 Cells , Hepacivirus/drug effects , Humans , Myxovirus Resistance Proteins/genetics , Protein Binding/drug effects , Vero Cells
13.
J Med Virol ; 92(9): 1649-1656, 2020 09.
Article in English | MEDLINE | ID: mdl-32239522

ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes the recent COVID-19 public health crisis. Bat is the widely believed original host of SARS-CoV-2. However, its intermediate host before transmitting to humans is not clear. Some studies proposed pangolin, snake, or turtle as the intermediate hosts. Angiotensin-converting enzyme 2 (ACE2) is the receptor for SARS-CoV-2, which determines the potential host range for SARS-CoV-2. On the basis of structural information of the complex of human ACE2 and SARS-CoV-2 receptor-binding domain (RBD), we analyzed the affinity to S protein of the 20 key residues in ACE2 from mammal, bird, turtle, and snake. Several ACE2 proteins from Primates, Bovidae, Cricetidae, and Cetacea maintained the majority of key residues in ACE2 for associating with SARS-CoV-2 RBD. The simulated structures indicated that ACE2 proteins from Bovidae and Cricetidae were able to associate with SARS-CoV-2 RBD. We found that nearly half of the key residues in turtle, snake, and bird were changed. The simulated structures showed several key contacts with SARS-CoV-2 RBD in turtle and snake ACE2 were abolished. This study demonstrated that neither snake nor turtle was the intermediate hosts for SARS-CoV-2, which further reinforced the concept that the reptiles are resistant against infection of coronavirus. This study suggested that Bovidae and Cricetidae should be included in the screening of intermediate hosts for SARS-CoV-2.


Subject(s)
Angiotensin-Converting Enzyme 2/metabolism , COVID-19/metabolism , COVID-19/virology , Receptors, Virus/metabolism , SARS-CoV-2/physiology , Spike Glycoprotein, Coronavirus/metabolism , Amino Acid Sequence , Angiotensin-Converting Enzyme 2/chemistry , Animals , Arvicolinae , Cattle , Humans , Models, Molecular , Multiprotein Complexes/chemistry , Multiprotein Complexes/metabolism , Protein Binding , Receptors, Virus/chemistry , Sequence Alignment , Spike Glycoprotein, Coronavirus/chemistry , Spike Glycoprotein, Coronavirus/genetics , Structure-Activity Relationship , Viral Tropism
14.
Microbiol Immunol ; 64(8): 574-577, 2020 Aug.
Article in English | MEDLINE | ID: mdl-32108375

ABSTRACT

Previously, it was found that the hepatitis C virus NS5A interacted with ACBD3 in a genotype-dependent manner. However, the region in NS5A responsible for association with ACBD3 is not clear. Domain I of NS5A was identified as critical for ACBD3 binding. By comparing the differences of amino acids in domain I from different genotypes of NS5A, it was found that key amino acids potentially corresponded to the affinity of the NS5A-ACBD3 interaction. The findings not only revealed that domain I of NS5A associates with ACBD3 but they also shed mechanistic light on how NS5A is associated with ACBD3 in a genotype-dependent manner.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Hepacivirus/genetics , Membrane Proteins/metabolism , Viral Nonstructural Proteins/genetics , Amino Acid Sequence , Cell Line , Genotype , HEK293 Cells , Hepatitis C/virology , Host Microbial Interactions , Humans , Protein Binding , Protein Conformation , Protein Domains , Viral Nonstructural Proteins/metabolism
15.
Biochem Biophys Res Commun ; 513(4): 919-924, 2019 06 11.
Article in English | MEDLINE | ID: mdl-31005258

ABSTRACT

Chikungunya virus (CHIKV) is a re-emerging mosquito-transmitted RNA virus causing joint and muscle pain. Although the protein-protein interactions (PPIs) between nonstructural proteins of CHIKV have been extensively established, the complete CHIKV intraviral interactome remains to be elucidated. In this study, we examined all possible CHIKV intraviral PPIs by immunoprecipitation and constructed the intraviral interactome of CHIKV. We reported 19 novel PPIs including the homo-oligomerization of TF. Disulfide bonds promoted the oligomerization of CHIKV TF protein. 2-BP, a palmitoylation inhibitor reduced the palmitoylation of TF and increased TF oligomerization. A quadruple mutant of Cys33, Cys35, Cys41, and Cys43 in TF blocked its palmitoylation and reduced oligomerization. Furthermore, we determined the association of TF with nsP1 and nsP3 in a palmitoylation-dependent manner. Construction of intraviral interactome of CHIKV provides the basis for further studying the function of CHIKV proteins.


Subject(s)
Chikungunya virus/metabolism , Protein Interaction Mapping/methods , Viral Proteins/metabolism , Immunoprecipitation , Lipoylation , Protein Multimerization , Viral Nonstructural Proteins/metabolism , Viral Structural Proteins/metabolism
16.
J Virol ; 91(18)2017 09 15.
Article in English | MEDLINE | ID: mdl-28679754

ABSTRACT

Claudin-1 is a hepatitis C virus (HCV) coreceptor required for viral entry. Although extensive studies have focused on claudin-1 as an anti-HCV target, little is known about how the level of claudin-1 at the cell surface is regulated by host vesicular transport. Here, we identified an interaction between claudin-1 and Sec24C, a cargo-sorting component of the coat protein complex II (COPII) vesicular transport system. By interacting with Sec24C through its C-terminal YV, claudin-1 is transported from the endoplasmic reticulum (ER) and is eventually targeted to the cell surface. Blocking COPII transport inhibits HCV entry by reducing the level of claudin-1 at the cell surface. These findings provide mechanistic insight into the role of COPII vesicular transport in HCV entry.IMPORTANCE Tight junction protein claudin-1 is one of the cellular receptors for hepatitis C virus, which infects 185 million people globally. Its cellular distribution plays important role in HCV entry; however, it is unclear how the localization of claudin-1 to the cell surface is controlled by host transport pathways. In this paper, we not only identified Sec24C as a key host factor for HCV entry but also uncovered a novel mechanism by which the COPII machinery transports claudin-1 to the cell surface. This mechanism might be extended to other claudins that contain a C-terminal YV or V motif.


Subject(s)
Claudin-1/metabolism , Hepacivirus/physiology , Receptors, Virus/metabolism , Vesicular Transport Proteins/metabolism , Virus Internalization , Cell Line , Hepatocytes/immunology , Hepatocytes/physiology , Humans , Protein Interaction Mapping , Protein Transport
17.
Biochem Biophys Res Commun ; 493(3): 1190-1196, 2017 11 25.
Article in English | MEDLINE | ID: mdl-28962860

ABSTRACT

The infection of CHIKV is associated with cellular membranes; however whether early secretory pathways are involved in CHIKV replication remains unclear. In the present study, we have provided initial evidences that CHIKV requires both COPI and COPII for its replication. Small interfering RNAs against COPI components, including coatomer, ARFs or GBF1, suppress CHIKV replication. Moreover, CHIKV infection is abolished by the presence of ARF1 inhibitor brefeldin A or GBF1 inhibitor golgicide A. In addition, perturbation of COPII by silencing key components of COPII pathways leads to a reduction in CHIKV replication. Collectively, these observations demonstrate the importance of functional secretory pathways in the infectivity of CHIKV.


Subject(s)
Chikungunya virus/physiology , Coat Protein Complex I/metabolism , Viral Proteins/metabolism , Virus Replication/physiology , ADP-Ribosylation Factor 1/genetics , ADP-Ribosylation Factor 1/metabolism , Brefeldin A/pharmacology , Chikungunya virus/pathogenicity , Coat Protein Complex I/genetics , Guanine Nucleotide Exchange Factors/genetics , Guanine Nucleotide Exchange Factors/metabolism , HeLa Cells , Humans , Pyridines/pharmacology , Quinolines/pharmacology , RNA, Small Interfering , Viral Proteins/genetics , Virus Replication/drug effects
18.
Virus Genes ; 53(6): 814-822, 2017 Dec.
Article in English | MEDLINE | ID: mdl-28815417

ABSTRACT

Identification of viral-host interacting proteins will contribute to understanding of how poxvirus exploits the host cellular machinery. The vaccinia virus gene K7R encodes a conserved protein K7 in most orthopoxviruses. To gain insight into the biology of K7, we investigated the cellular interactome of K7 by GST pulldown coupled with mass spectrometry. The top categories of identified proteins contained components of trafficking machineries. We selected key components of three transport machineries including coatomer, retromer, and CHEVI to further confirm their binding abilities to K7. Di-lysine motif of K7 is required for its interaction with coatomer, while C terminal leucines in K7 are critical for association of retromer. Our study uncovers the viral-host interactome of vaccinia K7 and reveals three host transport machineries as binding partners of K7, which might have important roles in poxvirus' life cycles.


Subject(s)
Host-Pathogen Interactions/physiology , Protein Binding/physiology , Vaccinia virus/metabolism , Viral Proteins/metabolism , Cell Line, Tumor , HeLa Cells , Humans , Vaccinia/metabolism , Vaccinia/virology
19.
Cell Mol Life Sci ; 73(4): 869-81, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26298293

ABSTRACT

Hepatitis C virus (HCV) has infected over 170 million people worldwide. Phosphatidylinositol 4-phosphate (PI4P) is the organelle-specific phosphoinositide enriched at sites of HCV replication. Whether retromer, a PI4P-related host transport machinery, unloads its cargo at HCV replication sites remains inconclusive. We sought to characterize the role of retromer in HCV replication. Here, we demonstrated the interaction between retromer subunit Vps35 and HCV NS5A protein by immunoprecipitation and GST pulldown. Vps35 colocalized with NS5A and PI4P in both OR6 replicon and JFH1 infected Huh 7.5.1 cells. HCV replication was inhibited upon silencing retromer subunits. CIMPR, a typical retromer cargo, participated in HCV replication. Our data suggest that retromer component Vps35 is recruited by NS5A to viral replication sites where PI4P unloads CIMPR. These findings demonstrate a dependence role of retromer in HCV replication and identify retromer as a potential therapeutic target against HCV.


Subject(s)
Hepacivirus/physiology , Hepatitis C/metabolism , Hepatitis C/virology , Phosphatidylinositol Phosphates/metabolism , Vesicular Transport Proteins/metabolism , Viral Nonstructural Proteins/metabolism , Virus Replication , Cell Line , Humans , Phosphatidylinositol Phosphates/analysis , Vesicular Transport Proteins/analysis , Viral Nonstructural Proteins/analysis
20.
Biochem Biophys Res Commun ; 475(1): 31-6, 2016 06 17.
Article in English | MEDLINE | ID: mdl-27157138

ABSTRACT

Lipid droplets are the place for HCV assembly and ADRP is an abundant lipid droplets-associated protein. However, little is known about the mechanisms how ADRP is involved in HCV life cycle. Here we demonstrate that activation of ARF1 dissociates ADRP from lipid droplets. A constitute active form of ARF1 (ARF1Q71I) promotes HCV assembly. We found that ADRP plays a positive role in HCV replication and a negative role in HCV assembly. Overexpression of ADRP increases the size of lipid droplets, while silencing ADRP reduces the size of lipid droplets. These findings provide new insight into the role of lipid droplets proteins in life cycle of HCV.


Subject(s)
ADP-Ribosylation Factor 1/metabolism , Hepacivirus/physiology , Hepatitis C/metabolism , Lipid Droplets/virology , Perilipin-2/metabolism , Virus Assembly , Cell Line , Hepatitis C/virology , Host-Pathogen Interactions , Humans , Lipid Droplets/metabolism , Virus Replication
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