Molecular Mechanisms of Parvovirus Gene Expression and Replication

Summary

Principal Investigator: Jianming Qiu
Abstract: DESCRIPTION (provided by applicant): Human parvovirus B19 (B19V) infection causes hydrops fetalis in pregnant women during the second- trimester. It also causes severe hematological diseases, including transient aplastic crisis in patients with a high turn-over rate f red blood cells;and chronic anemia in immunodeficient and immunocompromised patients which, in some cases, can be fatal. At present, no specific antiviral drugs or vaccines (that prevent B19V infection in high-risk groups) are available. B19V replication is highly restricted to human erythroid progenitor cells (EPCs) in the bone marrow and fetal liver. B19V infection-mediated hydrops fetalis and hematological disorders mainly result from the direct killing of the EPCs in which B19V replicates. Among DNA viruses, B19V has a unique feature in the processing of its precursor mRNA (pre-mRNA) in that all the viral mRNAs are alternatively processed from a single pre-mRNA. Alternative splicing of the B19V pre-mRNA, which is controlled by multiple splicing enhancers, plays a key role in regulating alternative polyadenylation of B19V mRNAs, and generates abundant small viral mRNAs for encoding two small non- structural viral proteins, i.e. 7.5-kDa and 11-kDa. The 11-kDa protein interacts with Grb2, a protein that links the signals mediated by the erythropoietin receptor to the Ras/MEK/ERK pathway in EPCs, and plays an important role in viral DNA replication. Importantly, B19V infection induces a DNA damage response (DDR). Activation of ATR and DNA-PKcs facilitates viral DNA replication. B19V does not use the host replication machinery to replicate its ssDNA genome;rather, it appears to induce a DDR and subsequently co-opts the host DNA repair mechanism to facilitate its own replication. Furthermore, B19V replication in EPCs is markedly increased under hypoxic conditions and is mediated via the down-regulation of MEK/ERK signaling. Over the past few years, we have established two experimental cell systems that remove two critical barriers to the study of B19V replication: an efficient system of productive B19V infection involving the ex vivo- expansion of EPCs under hypoxic conditions (which mimic the microenvironment of EPCs in human bone marrow and fetal liver), and a reverse genetics approach that involves transfection of the B19V dsDNA-form genome into UT7/Epo-S1 cell line cells cultured under hypoxic conditions. Using these two systems, as well as a newly-established in vitro assay to measure viral DNA replication, we will: i) determine the mechanisms underlying the alternative processing of B19V pre-mRNA;ii) elucidate the mechanisms by which the 11-kDa protein subverts MEK/ERK signaling to promote B19V replication;and iii) determine the mechanisms underlying DDR-facilitated B19V DNA replication. Our long-term goal is to identify the key molecular mechanisms underlying the alterative processing of the single promoter-transcribed parvoviral pre-mRNA, as well as parvovirus DNA replication, in a physiologically- relevant setting, i.e., EPCs ex vivo-expanded under hypoxic conditions for B19V in this application.
Funding Period: 2007-08-01 - 2018-04-30
more information: NIH RePORT

Top Publications

  1. pmc ELISAs using human bocavirus VP2 virus-like particles for detection of antibodies against HBoV
    Feng Lin
    Wenling Hospital of Wenzhou Medical College, Zhejiang, China
    J Virol Methods 149:110-7. 2008
  2. pmc Productive parvovirus B19 infection of primary human erythroid progenitor cells at hypoxia is regulated by STAT5A and MEK signaling but not HIFα
    Aaron Yun Chen
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, United States of America
    PLoS Pathog 7:e1002088. 2011
  3. pmc Molecular characterization of the newly identified human parvovirus 4 in the family Parvoviridae
    Sai Lou
    Department of Infectious Diseases, First Affiliated Hospital, School of Medicine, Xi an Jiaotong University, Xi an, China
    Virology 422:59-69. 2012
  4. pmc Internal polyadenylation of parvoviral precursor mRNA limits progeny virus production
    Qinfeng Huang
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas 66160, USA
    Virology 426:167-77. 2012
  5. pmc Human parvovirus B19 DNA replication induces a DNA damage response that is dispensable for cell cycle arrest at phase G2/M
    Sai Lou
    Department of Infectious Diseases, First Affiliated Hospital, School of Medicine, Xi an Jiaotong University, Xi an, China
    J Virol 86:10748-58. 2012
  6. pmc Establishment of a reverse genetics system for studying human bocavirus in human airway epithelia
    Qinfeng Huang
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA
    PLoS Pathog 8:e1002899. 2012
  7. pmc Structure of the NS1 protein N-terminal origin recognition/nickase domain from the emerging human bocavirus
    Sunil Kumar Tewary
    Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas, USA
    J Virol 87:11487-93. 2013
  8. pmc Human parvovirus B19 infection causes cell cycle arrest of human erythroid progenitors at late S phase that favors viral DNA replication
    Yong Luo
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA
    J Virol 87:12766-75. 2013
  9. pmc Human bocavirus 1 infects commercially available primary human airway epithelium cultures productively
    Xuefeng Deng
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS, USA
    J Virol Methods 195:112-9. 2014
  10. pmc The family Parvoviridae
    Susan F Cotmore
    Department of Laboratory Medicine, Yale University School of Medicine, New Haven, CT, USA
    Arch Virol 159:1239-47. 2014

Research Grants

  1. Viral Modulation of Genetic Stability
    Matthew D Weitzman; Fiscal Year: 2013
  2. Serotonin as a Regulator of Bone Mass Accrual: Basic and Clinical
    Gerard Karsenty; Fiscal Year: 2013
  3. DEVELOPMENTAL BIOLOGY OF HUMAN ERYTHROPOIESIS
    Stuart H Orkin; Fiscal Year: 2013
  4. B-cell Biology of Mucosal Immune Protection from SIV Challenge
    Eric Hunter; Fiscal Year: 2013
  5. PROTON RADIATION THERAPY RESEARCH
    Thomas F DeLaney; Fiscal Year: 2013
  6. Primary Immune Deficiency Treatment Consortium
    Morton Cowan; Fiscal Year: 2013
  7. The Virtual Physiological Rat Project
    Daniel A Beard; Fiscal Year: 2013
  8. ADULT LEUKEMIA RESEARCH CENTER
    Frederick Appelbaum; Fiscal Year: 2013
  9. Regulation of HSV-1 Gene Expression and Replication by Nuclear Hormone Receptors
    SHAOCHUNG VICTOR HSIA; Fiscal Year: 2013
  10. Rand Center of Excellence for the Study of Appropriateness of Care in CAM
    IAN DOUGLASS COULTER; Fiscal Year: 2013

Detail Information

Publications26

  1. pmc ELISAs using human bocavirus VP2 virus-like particles for detection of antibodies against HBoV
    Feng Lin
    Wenling Hospital of Wenzhou Medical College, Zhejiang, China
    J Virol Methods 149:110-7. 2008
    ..Thus, HBoV appears to be a common infection in children. The potential pathogenesis of this virus, especially its role in lower respiratory tract infections in children warrants further investigation...
  2. pmc Productive parvovirus B19 infection of primary human erythroid progenitor cells at hypoxia is regulated by STAT5A and MEK signaling but not HIFα
    Aaron Yun Chen
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, United States of America
    PLoS Pathog 7:e1002088. 2011
    ....
  3. pmc Molecular characterization of the newly identified human parvovirus 4 in the family Parvoviridae
    Sai Lou
    Department of Infectious Diseases, First Affiliated Hospital, School of Medicine, Xi an Jiaotong University, Xi an, China
    Virology 422:59-69. 2012
    ..Taken together, our characterization of the molecular features of PARV4 suggests that PARV4 represents a new genus in the family Parvoviridae...
  4. pmc Internal polyadenylation of parvoviral precursor mRNA limits progeny virus production
    Qinfeng Huang
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas 66160, USA
    Virology 426:167-77. 2012
    ....
  5. pmc Human parvovirus B19 DNA replication induces a DNA damage response that is dispensable for cell cycle arrest at phase G2/M
    Sai Lou
    Department of Infectious Diseases, First Affiliated Hospital, School of Medicine, Xi an Jiaotong University, Xi an, China
    J Virol 86:10748-58. 2012
    ..Taken together, the results suggest that the replication of the B19V genome is largely responsible for triggering a DDR, which does not perturb cell cycle progression at G(2)/M significantly, during B19V infection...
  6. pmc Establishment of a reverse genetics system for studying human bocavirus in human airway epithelia
    Qinfeng Huang
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA
    PLoS Pathog 8:e1002899. 2012
    ..Thus, we have established a reverse genetics system and generated the first cell line-based culture system for the study of HBoV1 infection, which will significantly advance the study of HBoV1 replication and pathogenesis...
  7. pmc Structure of the NS1 protein N-terminal origin recognition/nickase domain from the emerging human bocavirus
    Sunil Kumar Tewary
    Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas, USA
    J Virol 87:11487-93. 2013
    ....
  8. pmc Human parvovirus B19 infection causes cell cycle arrest of human erythroid progenitors at late S phase that favors viral DNA replication
    Yong Luo
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA
    J Virol 87:12766-75. 2013
    ..Taken together, our results confirmed that B19V infection triggers late S phase arrest, which presumably provides cellular S phase factors for viral DNA replication. ..
  9. pmc Human bocavirus 1 infects commercially available primary human airway epithelium cultures productively
    Xuefeng Deng
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS, USA
    J Virol Methods 195:112-9. 2014
    ..6 cm(2)). This study is the first to use commercial sources of HAE cultures for HBoV1 infection. The availability of these cultures will enable a wide range of laboratories to study HBoV1 infection...
  10. pmc The family Parvoviridae
    Susan F Cotmore
    Department of Laboratory Medicine, Yale University School of Medicine, New Haven, CT, USA
    Arch Virol 159:1239-47. 2014
    ..Also, affixes will be included in the names of genera to clarify subfamily affiliation and reduce the ambiguity that results from the vernacular use of "parvovirus" and "densovirus" to denote multiple taxon levels. ..
  11. pmc The human parvovirus B19 non-structural protein 1 N-terminal domain specifically binds to the origin of replication in the viral DNA
    Sunil Kumar Tewary
    Department of Molecular Biosciences, University of Kansas, 1200 Sunnyside Avenue, Lawrence, KS 66045, USA
    Virology 449:297-303. 2014
    ..Such a specialized nucleoprotein complex may enable NS1 to nick the terminal resolution site and separate DNA strands during replication. ..
  12. pmc Parvovirus B19 infection of human primary erythroid progenitor cells triggers ATR-Chk1 signaling, which promotes B19 virus replication
    Yong Luo
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USA
    J Virol 85:8046-55. 2011
    ..Notably, the cell cycle arrest characteristic of B19V infection was not rescued by interference with the activity of any of the three repair pathway kinases...
  13. pmc Internal polyadenylation of the parvovirus B19 precursor mRNA is regulated by alternative splicing
    Wuxiang Guan
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas 66160, USA
    J Biol Chem 286:24793-805. 2011
    ..Thus, our study reveals the mechanism by which alternative splicing coordinates alternative polyadenylation to generate full-length B19V mRNA transcripts at levels sufficient to support productive B19V infection...
  14. pmc Block to the production of full-length B19 virus transcripts by internal polyadenylation is overcome by replication of the viral genome
    Wuxiang Guan
    University of Kansas Medical Center, MS3029, 3901 Rainbow Blvd, Kansas City, KS 66160, USA
    J Virol 82:9951-63. 2008
    ..Furthermore, we show that polyadenylation of B19 viral RNA at (pA)p likely competes with splicing at the second intron. Thus, we conclude that replication of the B19 virus genome is the primary limiting step governing B19 virus tropism...
  15. pmc Molecular characterization of infectious clones of the minute virus of canines reveals unique features of bocaviruses
    Yuning Sun
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Mail Stop 3029, 3901 Rainbow Blvd, Kansas City, KS 66160, USA
    J Virol 83:3956-67. 2009
    ..Thus, our studies revealed important information about the genus Bocavirus that may eventually help us to clone the human bocavirus and study its pathogenesis...
  16. pmc The genome of human parvovirus b19 can replicate in nonpermissive cells with the help of adenovirus genes and produces infectious virus
    Wuxiang Guan
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, USA
    J Virol 83:9541-53. 2009
    ..Our results have important implications for our understanding of native B19V infection...
  17. pmc Molecular characterization of human parvovirus B19 genotypes 2 and 3
    Zhaojun Chen
    Department of Clinical Laboratory, the Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang Province, China
    Virology 394:276-85. 2009
    ..Similar to the proapoptotic nature of the prototype B19V large non-structural protein (NS1), the A6 and V9 NS1 proteins also are potent inducers of apoptosis in B19V-permissive cells...
  18. pmc The small 11 kDa nonstructural protein of human parvovirus B19 plays a key role in inducing apoptosis during B19 virus infection of primary erythroid progenitor cells
    Aaron Yun Chen
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USA
    Blood 115:1070-80. 2010
    ..Taken together, these results demonstrate that the 11 kDa protein contributes to erythroid progenitor cell death during B19V infection...
  19. pmc Bocavirus infection induces mitochondrion-mediated apoptosis and cell cycle arrest at G2/M phase
    Aaron Yun Chen
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Mail Stop 3029, 3901 Rainbow Blvd, Kansas City, KS 66160, USA
    J Virol 84:5615-26. 2010
    ..Our results may shed light on the molecular pathogenesis of Bocavirus infection in general...
  20. pmc Characterization of the gene expression profile of human bocavirus
    Aaron Yun Chen
    Department of Microbiology, University of Kansas Medical Center, Kansas City, KS, USA
    Virology 403:145-54. 2010
    ..Moreover, our results showed that, unlike human parvovirus B19 infection, expression of the HBoV1 proteins only does not induce cell cycle arrest and apoptosis of A549 cells...
  21. pmc Role of erythropoietin receptor signaling in parvovirus B19 replication in human erythroid progenitor cells
    Aaron Yun Chen
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas 66160, USA
    J Virol 84:12385-96. 2010
    ....
  22. pmc Chipmunk parvovirus is distinct from members in the genus Erythrovirus of the family Parvoviridae
    Zhaojun Chen
    Department of Clinical Laboratory, the Affiliated Hospital of Hangzhou Normal University, Hangzhou, China
    PLoS ONE 5:e15113. 2010
    ..Thus, we conclude that ChpPV may represent a new genus in the family Parvoviridae...
  23. pmc Inclusion of the central exon of parvovirus B19 precursor mRNA is determined by multiple splicing enhancers in both the exon and the downstream intron
    Wuxiang Guan
    Department of Microbiology, Molecular Genetics and Immunology, Kansas City, KS 66160, USA
    J Virol 85:2463-8. 2011
    ....
  24. pmc Molecular characterization of the small nonstructural proteins of parvovirus Aleutian mink disease virus (AMDV) during infection
    Qinfeng Huang
    Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, United States
    Virology 452:23-31. 2014
    ..More importantly, we provided evidence that both NS2 and NS3 are necessary for AMDV replication. ..

Research Grants30

  1. Viral Modulation of Genetic Stability
    Matthew D Weitzman; Fiscal Year: 2013
    ..These studies will elucidate the biological relevance of the interactions between cellular repair factors and viral proteins, and will have broader implications for our understanding of cellular DNA damage responses. ..
  2. Serotonin as a Regulator of Bone Mass Accrual: Basic and Clinical
    Gerard Karsenty; Fiscal Year: 2013
    ..Together our studies should provide important and novel insights in the genetic and molecular control of bone remodeling as well as in the pathogenesis and treatment of osteoporosis, a major disease of aging. ..
  3. DEVELOPMENTAL BIOLOGY OF HUMAN ERYTHROPOIESIS
    Stuart H Orkin; Fiscal Year: 2013
    ..abstract_text> ..
  4. B-cell Biology of Mucosal Immune Protection from SIV Challenge
    Eric Hunter; Fiscal Year: 2013
    ....
  5. PROTON RADIATION THERAPY RESEARCH
    Thomas F DeLaney; Fiscal Year: 2013
    ..abstract_text> ..
  6. Primary Immune Deficiency Treatment Consortium
    Morton Cowan; Fiscal Year: 2013
    ..These studies will resolve critical questions concerning HCT for these disorders and form the basis for future prospective clinical trials. ..
  7. The Virtual Physiological Rat Project
    Daniel A Beard; Fiscal Year: 2013
    ..This proposal targets the grand challenge of understanding complex multi-faceted disease phenotypes through experiments and simulations that capture the complex genotype-environment-phenotype relationship. ..
  8. ADULT LEUKEMIA RESEARCH CENTER
    Frederick Appelbaum; Fiscal Year: 2013
    ..existence of a large group of investigators all focused on the general topic of HCT;(2) strong preclinical research programs at our Center in support of this topic;and, (3) treatment of >500 transplant patients at our Center each year ..
  9. Regulation of HSV-1 Gene Expression and Replication by Nuclear Hormone Receptors
    SHAOCHUNG VICTOR HSIA; Fiscal Year: 2013
    ..The long-term goal is to identify the regulatory mechanisms to assist in the development of novel therapeutic protocols for better treatments. ..
  10. Rand Center of Excellence for the Study of Appropriateness of Care in CAM
    IAN DOUGLASS COULTER; Fiscal Year: 2013
    ..From these data are abstracted about the treatment given for CCP and the number of times M/M is done for CCP is obtained. We will also interview a sample of patients attending the clinics and do a follow study for their outcomes. ..
  11. Using Chemical Biology to Interfere with the Influenza Virus Life Cycle
    Michael G Roth; Fiscal Year: 2013
    ..In sum, these studies will likely reveal novel leads for antiviral therapies as well as provide information on novel mechanisms of viral-host interactions and pathways. ..
  12. Protein homeostasis mechanisms underlying enterovirus replication and evolution
    Raul Andino; Fiscal Year: 2013
    ..Core A: Administrative Core;and Core B: "High-throughput functional genomics and proteomics core. ..
  13. Immunobioogy for Marrow Allografts for Leukemia
    RICHARD JOHN O'REILLY; Fiscal Year: 2013
    ..The 3 cores include: Core A which provides all patient samples and evaluate grafts pre and post HSCT, Core B Biostatistics and Core C administrative support and oversight. ..
  14. Middle Atlantic Regional Center for Excellence for Biodefense and Emerging Infect
    MYRON MAX LEVINE; Fiscal Year: 2013
    ..abstract_text> ..
  15. Northeast Biodefense Center
    W Ian Lipkin; Fiscal Year: 2013
    ..As a Center based in a School of Public Health and a State Department of Health, the NBC has a firm commitment to and practical understanding of Emergency Preparedness. ..
  16. HORMONAL REGULATION OF BLOOD PRESSURE
    Michal Laniado Schwartzman; Fiscal Year: 2013
    ..ular tone, in the pathophysiology of hypertension and cardiovascular disease. ..
  17. Arterial Dysfunction: Basic and Clinical Mechanisms
    Thomas Michel; Fiscal Year: 2013
    ..Gladyshev. P. Libby directs the Redox Biomarkers Core;metabolic characterizations of mouse models studied in this Program will take place at the Yale Mouse Metabolic Phenotyping Center, led by G. Shulman. ..
  18. Enhancing radiation and cisplatin HNSCC cell killing by inhibiting mitochondrial
    Lynn Harrison; Fiscal Year: 2013
    ..Specific Aim 2 ..
  19. Host-tumor cell interaction in myeloma: therapeutic applications
    Kenneth C Anderson; Fiscal Year: 2013
    ..Ultimately, our goal is to validate MM-host cell interactions as a target for novel therapeutics to improve patient outcome in MM. ..
  20. Pacific Southwest RCE for Biodefense &Emerging Infectious Diseases Research
    Alan G Barbour; Fiscal Year: 2013
    ..abstract_text> ..
  21. STEM CELL GENE THERAPY FOR HEMOGLOBINOPATHIES
    George Stamatoyannopoulos; Fiscal Year: 2013
    ..The focus of this Program Project, Gene Therapy, can provide a new paradigm for the treatment of these hemoglobinopathies as well as for other blood diseases. (End of Abstract) ..
  22. Rocky Mountain Regional Center of Excellence or Biodefense and Emerging Infectiou
    John T Belisle; Fiscal Year: 2013
    ..abstract_text> ..
  23. VACCINE INDUCED IMMUNITY IN THE YOUNG AND AGED
    Rafi Ahmed; Fiscal Year: 2013
    ....
  24. Molecular Modulation of Virus Capsid Assembly
    Adam Zlotnick; Fiscal Year: 2013
    ..These independent aims will move assembly effectors from the subject of purely academic investigation towards clinical application. ..