Analysis of EV71 infection progression using triple‐SILAC‐based proteomics approach

Enterovirus 71 (EV71), a member of Picornaviridae, causes severe neurological and systemic illness in children. To better understand the virus–host cell interactions, we performed a triple‐SILAC‐based quantitative proteomics study monitoring host cell proteome changes after EV71 infection. Based on the quantitative data for more than 4100 proteins, ∼17% of the proteins were found as significantly changed (p<0.01) at either 8 or 20 hours post infection. Five biological processes and seven protein classes showed significant differences. Functional screening of nine regulated proteins discovered the regulatory role of CHCH2, a mitochondrial protein known as a transcriptional activator for cytochrome c oxidase, in EV71 replication. Further studies showed that CHCH2 served as a negative regulator of innate immune responses. All MS data have been deposited in the ProteomeXchange with identifier PXD002483 (http://proteomecentral.proteomexchange.org/dataset/PXD002483).

[1]  M. Katze,et al.  A proteomic glimpse into the initial global epigenetic changes during HIV infection , 2014, Proteomics.

[2]  R. Springett,et al.  MNRR1 (formerly CHCHD2) is a bi-organellar regulator of mitochondrial metabolism. , 2015, Mitochondrion.

[3]  R. Kuo,et al.  MDA5 Plays a Crucial Role in Enterovirus 71 RNA-Mediated IRF3 Activation , 2013, PloS one.

[4]  H. Shu,et al.  FoxO1 Negatively Regulates Cellular Antiviral Response by Promoting Degradation of IRF3* , 2013, The Journal of Biological Chemistry.

[5]  Shih-Cheng Chang,et al.  Viral and host proteins involved in picornavirus life cycle , 2009, Journal of Biomedical Science.

[6]  H. Gendelman,et al.  Functional Proteomic Analysis for Regulatory T Cell Surveillance of the HIV-1-Infected Macrophage , 2010, Journal of proteome research.

[7]  M. Stinski,et al.  Interaction Network of Proteins Associated with Human Cytomegalovirus IE2-p86 Protein during Infection: A Proteomic Analysis , 2013, PloS one.

[8]  P. Tien,et al.  Cell Surface Vimentin Is an Attachment Receptor for Enterovirus 71 , 2014, Journal of Virology.

[9]  Andrew R. Jones,et al.  ProteomeXchange provides globally co-ordinated proteomics data submission and dissemination , 2014, Nature Biotechnology.

[10]  S. Tikoo,et al.  Viruses as Modulators of Mitochondrial Functions , 2013, Advances in virology.

[11]  P. McMinn Recent advances in the molecular epidemiology and control of human enterovirus 71 infection. , 2012, Current opinion in virology.

[12]  Kuender D Yang,et al.  Sialylated glycans as receptor and inhibitor of enterovirus 71 infection to DLD-1 intestinal cells , 2009, Virology Journal.

[13]  Yi Zhang,et al.  Role of Bmi-1 and Ring1A in H2A ubiquitylation and Hox gene silencing. , 2005, Molecular cell.

[14]  Jincheng Li,et al.  Mitochondrial fission leads to Smac/DIABLO release quenched by ARC , 2010, Apoptosis.

[15]  R. Finley,et al.  Oxygen-dependent expression of cytochrome c oxidase subunit 4-2 gene expression is mediated by transcription factors RBPJ, CXXC5 and CHCHD2 , 2013, Nucleic acids research.

[16]  Peter A. DiMaggio,et al.  Quantitative Proteomic Discovery of Dynamic Epigenome Changes that Control Human Cytomegalovirus (HCMV) Infection * , 2014, Molecular & Cellular Proteomics.

[17]  M. Ho,et al.  Annexin II Binds to Capsid Protein VP1 of Enterovirus 71 and Enhances Viral Infectivity , 2011, Journal of Virology.

[18]  Jianmin Wang,et al.  Quinacrine Impairs Enterovirus 71 RNA Replication by Preventing Binding of Polypyrimidine-Tract Binding Protein with Internal Ribosome Entry Sites , 2013, PloS one.

[19]  N. Altan-Bonnet,et al.  Enteroviruses harness the cellular endocytic machinery to remodel the host cell cholesterol landscape for effective viral replication. , 2013, Cell host & microbe.

[20]  E. Emmott,et al.  Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics , 2014, Journal of visualized experiments : JoVE.

[21]  J. Barr,et al.  A quantitative proteomic analysis of lung epithelial (A549) cells infected with 2009 pandemic influenza A virus using stable isotope labelling with amino acids in cell culture , 2012, Proteomics.

[22]  H. Shu,et al.  E3 ligase WWP2 negatively regulates TLR3-mediated innate immune response by targeting TRIF for ubiquitination and degradation , 2013, Proceedings of the National Academy of Sciences.

[23]  S. Koike,et al.  Scavenger receptor B2 is a cellular receptor for enterovirus 71 , 2009, Nature Medicine.

[24]  Qibin Zhang,et al.  Temporal Proteome and Lipidome Profiles Reveal Hepatitis C Virus-Associated Reprogramming of Hepatocellular Metabolism and Bioenergetics , 2010, PLoS pathogens.

[25]  I. Cristea,et al.  A Proteomic Perspective of Inbuilt Viral Protein Regulation: pUL46 Tegument Protein is Targeted for Degradation by ICP0 during Herpes Simplex Virus Type 1 Infection* , 2013, Molecular & Cellular Proteomics.

[26]  R. Cortese,et al.  Cell Entry of Hepatitis C Virus Requires a Set of Co-receptors That Include the CD81 Tetraspanin and the SR-B1 Scavenger Receptor* , 2003, Journal of Biological Chemistry.

[27]  T. Solomon,et al.  Clinical features, diagnosis, and management of enterovirus 71 , 2010, The Lancet Neurology.

[28]  Jau-Song Yu,et al.  Proteomics analysis of EV71-infected cells reveals the involvement of host protein NEDD4L in EV71 replication. , 2015, Journal of proteome research.

[29]  T. Dobner,et al.  Human Pathogens and the Host Cell SUMOylation System , 2011, Journal of Virology.

[30]  M. Shimojima,et al.  Human P-selectin glycoprotein ligand-1 is a functional receptor for enterovirus 71 , 2009, Nature Medicine.

[31]  R. Guerra-Sá,et al.  Coxsackievirus B5 induced apoptosis of HeLa cells: effects on p53 and SUMO. , 2010, Virology.

[32]  Paul Ahlquist,et al.  Host Factors in Positive-Strand RNA Virus Genome Replication , 2003, Journal of Virology.

[33]  Jun Yu,et al.  Enterovirus 71 Disrupts Interferon Signaling by Reducing the Level of Interferon Receptor 1 , 2012, Journal of Virology.

[34]  H. Shu,et al.  ECSIT Bridges RIG-I-Like Receptors to VISA in Signaling Events of Innate Antiviral Responses , 2014, Journal of Innate Immunity.

[35]  M. Shipp,et al.  BBAP monoubiquitylates histone H4 at lysine 91 and selectively modulates the DNA damage response. , 2009, Molecular cell.

[36]  Robert E. Lewis,et al.  Ras regulates assembly of mitogenic signalling complexes through the effector protein IMP , 2004, Nature.

[37]  Z. Ronai,et al.  SUMO-1 modification of Mdm2 prevents its self-ubiquitination and increases Mdm2 ability to ubiquitinate p53. , 2000, Cell.

[38]  A. Burlingame,et al.  Quantitative proteomic analysis of Huh-7 cells infected with Dengue virus by label-free LC-MS. , 2014, Journal of proteomics.

[39]  C. Rogier,et al.  Identification of Cellular Proteome Modifications in Response to West Nile Virus Infection* , 2009, Molecular & Cellular Proteomics.

[40]  K. Nakayama,et al.  Nedd4-interacting protein 2, a short half-life membrane protein degraded in lysosomes, negatively controls down-regulation of connexin43. , 2010, Biological & pharmaceutical bulletin.

[41]  H. Shih,et al.  Sumoylation-promoted Enterovirus 71 3C Degradation Correlates with a Reduction in Viral Replication and Cell Apoptosis* , 2011, The Journal of Biological Chemistry.

[42]  V. Stollar,et al.  Host Factors in Enterovirus 71 Replication , 2011, Journal of Virology.

[43]  I. Sam,et al.  Enterovirus 71 Uses Cell Surface Heparan Sulfate Glycosaminoglycan as an Attachment Receptor , 2012, Journal of Virology.

[44]  M. Katze,et al.  Comprehensive Proteomic Analysis of Influenza Virus Polymerase Complex Reveals a Novel Association with Mitochondrial Proteins and RNA Polymerase Accessory Factors , 2011, Journal of Virology.

[45]  S. Hanash,et al.  Modification of Host Lipid Raft Proteome upon Hepatitis C Virus Replication*S , 2006, Molecular & Cellular Proteomics.

[46]  Roger A. Moore,et al.  Activation of the innate signaling molecule MAVS by bunyavirus infection upregulates the adaptor protein SARM1, leading to neuronal death. , 2013, Immunity.

[47]  E. H. Lennette,et al.  An apparently new enterovirus isolated from patients with disease of the central nervous system. , 1974, The Journal of infectious diseases.

[48]  P. Hu,et al.  The role of von Willebrand factor as a biomarker of tumor development in hepatitis B virus-associated human hepatocellular carcinoma: a quantitative proteomic based study. , 2014, Journal of proteomics.

[49]  A. Halayko,et al.  Response of Primary Human Airway Epithelial Cells to Influenza Infection: A Quantitative Proteomic Study , 2012, Journal of proteome research.

[50]  R. Liu,et al.  Comprehensive proteomic analysis of host cell lipid rafts modified by HBV infection. , 2012, Journal of proteomics.

[51]  Tom Solomon,et al.  Virology, epidemiology, pathogenesis, and control of enterovirus 71. , 2010, The Lancet. Infectious diseases.

[52]  Mei-Ling Li,et al.  Far upstream element binding protein 2 interacts with enterovirus 71 internal ribosomal entry site and negatively regulates viral translation , 2008, Nucleic acids research.

[53]  J. Sung,et al.  Glucose-regulated Protein 78 Is an Intracellular Antiviral Factor against Hepatitis B Virus , 2009, Molecular & Cellular Proteomics.

[54]  Kuo-Feng Weng,et al.  Enterovirus 71 3C Protease Cleaves a Novel Target CstF-64 and Inhibits Cellular Polyadenylation , 2009, PLoS pathogens.

[55]  H. Shimizu,et al.  Cellular Receptors for Human Enterovirus Species A , 2012, Front. Microbio..

[56]  Hui Zhao,et al.  Global Transcriptomic Analysis of Human Neuroblastoma Cells in Response to Enterovirus Type 71 Infection , 2013, PloS one.

[57]  Gengfu Xiao,et al.  Identification of host proteins involved in Japanese encephalitis virus infection by quantitative proteomics analysis. , 2013, Journal of proteome research.

[58]  G. Panayotou,et al.  Comparative proteomic analysis implicates COMMD proteins as Epstein-Barr virus targets in the BL41 Burkitt's lymphoma cell line. , 2011, Journal of proteome research.

[59]  J. Horng,et al.  Enterovirus 71 Infection Cleaves a Negative Regulator for Viral Internal Ribosomal Entry Site-Driven Translation , 2013, Journal of Virology.

[60]  Anushya Muruganujan,et al.  Large-scale gene function analysis with the PANTHER classification system , 2013, Nature Protocols.

[61]  G. Multhaup,et al.  Foot-and-mouth disease virus protease 3C induces specific proteolytic cleavage of host cell histone H3 , 1990, Journal of virology.

[62]  Q. Jin,et al.  Enterovirus 71 Protease 2Apro Targets MAVS to Inhibit Anti-Viral Type I Interferon Responses , 2013, PLoS pathogens.

[63]  Z. Ronai,et al.  SUMO-1 Modification of Mdm2 Prevents Its Self-Ubiquitination and Increases Mdm2 Ability to Ubiquitinate p53 , 2000, Cell.

[64]  C. Ahlm,et al.  The Rift Valley Fever virus protein NSm and putative cellular protein interactions , 2012, Virology Journal.

[65]  V. Chow,et al.  Transcriptomic and proteomic analyses of rhabdomyosarcoma cells reveal differential cellular gene expression in response to enterovirus 71 infection , 2005, Cellular microbiology.

[66]  P. D. Nagy,et al.  The dependence of viral RNA replication on co-opted host factors , 2011, Nature Reviews Microbiology.

[67]  Yuquan Wei,et al.  An integrated proteomics and bioinformatics analyses of hepatitis B virus X interacting proteins and identification of a novel interactor apoA-I. , 2013, Journal of proteomics.

[68]  B. Semler,et al.  Inhibition of Poliovirus-Induced Cleavage of Cellular Protein PCBP2 Reduces the Levels of Viral RNA Replication , 2013, Journal of Virology.

[69]  Brian D. Peyser,et al.  Multi-Faceted Proteomic Characterization of Host Protein Complement of Rift Valley Fever Virus Virions and Identification of Specific Heat Shock Proteins, Including HSP90, as Important Viral Host Factors , 2014, PloS one.

[70]  Xuejun Jin,et al.  An Atypical E3 Ligase Zinc Finger Protein 91 Stabilizes and Activates NF-κB-inducing Kinase via Lys63-linked Ubiquitination* , 2010, The Journal of Biological Chemistry.

[71]  Jia Jun Lee,et al.  Comparative proteome analyses of host protein expression in response to Enterovirus 71 and Coxsackievirus A16 infections. , 2011, Journal of proteomics.

[72]  R. Cortese,et al.  The human scavenger receptor class B type I is a novel candidate receptor for the hepatitis C virus , 2002, The EMBO journal.

[73]  Q. Jin,et al.  The 3C Protein of Enterovirus 71 Inhibits Retinoid Acid-Inducible Gene I-Mediated Interferon Regulatory Factor 3 Activation and Type I Interferon Responses , 2010, Journal of Virology.

[74]  C. Boutell,et al.  Proteomic analysis of cells in the early stages of herpes simplex virus type‐1 infection reveals widespread changes in the host cell proteome , 2009, Proteomics.

[75]  J. Horng,et al.  Heterogeneous nuclear ribonuclear protein K interacts with the enterovirus 71 5' untranslated region and participates in virus replication. , 2008, The Journal of general virology.

[76]  Peng-Nien Huang,et al.  Update on enterovirus 71 infection. , 2014, Current opinion in virology.

[77]  Cameron J. Schweitzer,et al.  Proteomic analysis of early HIV-1 nucleoprotein complexes. , 2013, Journal of proteome research.

[78]  C. Coyne,et al.  Mechanisms of MAVS regulation at the mitochondrial membrane. , 2013, Journal of molecular biology.

[79]  A. Sparks,et al.  Identification of Novel Human WW Domain-containing Proteins by Cloning of Ligand Targets* , 1997, The Journal of Biological Chemistry.

[80]  T. Aittokallio,et al.  Quantitative Subcellular Proteome and Secretome Profiling of Influenza A Virus-Infected Human Primary Macrophages , 2011, PLoS pathogens.

[81]  C. Claus,et al.  A renewed focus on the interplay between viruses and mitochondrial metabolism , 2013, Archives of Virology.

[82]  P. Timms,et al.  Elucidating the host–pathogen interaction between human colorectal cells and invading Enterovirus 71 using transcriptomics profiling , 2014, FEBS open bio.

[83]  Zhijian J. Chen,et al.  Identification and Characterization of MAVS, a Mitochondrial Antiviral Signaling Protein that Activates NF-κB and IRF3 , 2005, Cell.