MECHANISMS OF HUMAN PAPILLOMAVIRUS DNA REPLICATION

Summary

Principal Investigator: LOUISE CHOW
Affiliation: University of Alabama at Birmingham
Country: USA
Abstract: Human papillomaviruses (HPVs) comprise an extended family of medically important human pathogens. These small DNA viruses establish persistent infections of squamous epithelia, typically causing benign hyperproliferative lesions. Infections by higher risk genotypes can progress to dysplasias and carcinomas, notably cervical and penile cancers. The double-stranded, circular genome replicates as extrachromosomal plasmids at low copy number in cycling basal keratinocytes. Productive amplification takes place only in differentiating spinous cells. We and others developed transient and cell-free systems to study the mechanisms of HPV DNA replication and showed that replication requires an origin of replication (ori), the HPV ori recognition protein E2, the viral replicative helicase E1, the cellular DNA replication machinery and cyclin/cdk complexes and that the E1 activity is regulated by multiple kinases. Our preliminary results reveal that a number of cellular proteins known to control chromosomal DNA replication also regulate HPV replication in the cell-free replication system, including Cdt1, geminin, TopBP1, and p53. Geminin inhibits Cdt1, which is necessary for loading the cellular replicative helicase MCM complex onto the cellular chromatin. Our results implicate MCM in controlling HPV replication when E1 concentration is low. TopBP1 is involved in both cellular DNA replication and repair. It interacts with DNA polymerase epsilon, a polymerase with critical but yet-to-be elucidated functions. TopBP1 has been reported to interact with HPV16 E2 and to stimulate transient HPV replication. We now show that TopBP1 enhances HPV cell-free replication. p53 is targeted by HPV E6 protein for inactivation and is known to bind E2 and inhibit viral transient amplification replication. We demonstrate that p53 inhibits HPV replication in the presence or absence of E2, indicative of additional mechanisms. We suggest that, in each case, the cell-free system provides an excellent opportunity to elucidate the mechanisms of regulation for both viral and cellular DNA replication. We propose four specific aims. 1. To test a new model for HPV DNA replication and regulation. This model would account for the maintenance mode and the amplification mode of viral DNA replication in infected tissues. 2. To determine the roles of TopBP1 and DNA polymerase epsilon in HPV DNA replication. 3. To test our hypothesis that p53 inhibition of HPV DNA replication is mediated at least in part by interfering with the activities of recombination proteins that have been hypothesized to be involved in chromosomal DNA replication. 4. To elucidate how E1 phosphorylation modulates its replication functions, a continuation of present research.
Funding Period: 1999-04-01 - 2010-03-31
more information: NIH RePORT

Top Publications

  1. ncbi Simultaneous in situ detection of RNA, DNA, and protein using tyramide-coupled immunofluorescence
    Brian A Van Tine
    Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA
    Methods Mol Biol 292:215-30. 2005
  2. pmc Dynamic localization of the human papillomavirus type 11 origin binding protein E2 through mitosis while in association with the spindle apparatus
    Luan D Dao
    Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294 0005, USA
    J Virol 80:4792-800. 2006
  3. pmc Conditionally activated E7 proteins of high-risk and low-risk human papillomaviruses induce S phase in postmitotic, differentiated human keratinocytes
    N Sanjib Banerjee
    Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294 0005, USA
    J Virol 80:6517-24. 2006
  4. pmc microRNAs are biomarkers of oncogenic human papillomavirus infections
    Xiaohong Wang
    Tumor Virus RNA Biology Section, Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892
    Proc Natl Acad Sci U S A 111:4262-7. 2014
  5. pmc Phylogenetic considerations in designing a broadly protective multimeric L2 vaccine
    Subhashini Jagu
    Department of Pathology, Johns Hopkins University, Baltimore, Maryland, USA
    J Virol 87:6127-36. 2013
  6. pmc HPV-18 E6 mutants reveal p53 modulation of viral DNA amplification in organotypic cultures
    Eun Young Kho
    Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294 0005, USA
    Proc Natl Acad Sci U S A 110:7542-9. 2013
  7. pmc Ubp-M serine 552 phosphorylation by cyclin-dependent kinase 1 regulates cell cycle progression
    Yang Xu
    Department of Biochemistry and Molecular Genetics University of Alabama at Birmingham Birmingham, AL USA State Key Laboratory of Cellular and Molecular Biology College of Life Science Hebei Normal University Shijiazhuang, China
    Cell Cycle 12:3219-27. 2013
  8. pmc Comprehensive control of human papillomavirus infections and related diseases
    F Xavier Bosch
    Cancer Epidemiology Research Program CERP, Institut Català d Oncologia Catalan Institute of Oncology ICO, IDIBELL, L Hospitalet de Llobregat Barcelona, Spain Electronic address
    Vaccine 31:I1-31. 2013
  9. ncbi Comprehensive control of human papillomavirus infections and related diseases
    F Xavier Bosch
    Cancer Epidemiology Research Program CERP, Institut Català d Oncologia Catalan Institute of Oncology ICO, IDIBELL, L Hospitalet de Llobregat Barcelona, Spain Electronic address
    Vaccine 31:F1-31. 2013
  10. ncbi Comprehensive control of human papillomavirus infections and related diseases
    F Xavier Bosch
    Cancer Epidemiology Research Program CERP, Institut Català d Oncologia Catalan Institute of Oncology ICO, IDIBELL, L Hospitalet de Llobregat Barcelona, Spain Electronic address
    Vaccine 31:G1-G31. 2013

Scientific Experts

  • LOUISE CHOW
  • Sarah M Bean
  • Sherry S Collawn
  • Florianne E Henken
  • Thomas R Broker
  • F Xavier Bosch
  • Hsu Kun Wang
  • Xiaohong Wang
  • John T Schiller
  • N Sanjib Banerjee
  • Nicholas J Genovese
  • Marc Steben
  • Philip E Castle
  • Eduardo L Franco
  • Anna Barbara Moscicki
  • Maura L Gillison
  • Rengaswamy Sankaranarayanan
  • David Forman
  • Mario Poljak
  • William A Fisher
  • Lynette A Denny
  • Lauri E Markowitz
  • Ginesa Albero
  • Maria Brotons
  • Marc Arbyn
  • Margaret Stanley
  • Jack Cuzick
  • Chris J L M Meijer
  • Laia Alemany
  • Mark Schiffman
  • Karen Canfell
  • Mireia Diaz
  • Peter L Stern
  • Mark A Kane
  • Xavier Castellsague
  • John Doorbar
  • Jane J Kim
  • Jei Hwa Yu
  • Craig Meyers
  • Zhi Ming Zheng
  • Yang Xu
  • Silvia De Sanjose
  • Subhashini Jagu
  • Eun Young Kho
  • Brian A Van Tine
  • Silvia de Sanjose
  • Jeonggu Sim
  • Biing Yuan Lin
  • Luan D Dao
  • Markus Hafner
  • Nilam Sanjib Banerjee
  • Yang Li
  • Daniel Briskin
  • Thomas Tuschl
  • Shuang Tang
  • Xing Xie
  • Archer D Smith
  • Sudha V Chivukula
  • David Schneider
  • Huirong Yang
  • MATTHEW RENFROW
  • Hengbin Wang
  • Harold Kleanthous
  • Warner K Huh
  • Kilvani S Jaganathan
  • Douglas R Lowy
  • Kirill Kalnin
  • Heui Yun Joo
  • Richard B S Roden
  • Kihyuck Kwak
  • Chenguang Wang
  • Janet S Rader
  • Aaron A Duffy
  • Nilam S Banerjee
  • J Philip McCoy
  • Jack Griffith
  • Sezgin Ozgur
  • Wentao Deng
  • Shwu Yuan Wu
  • Cheng Ming Chiang
  • Aaron Duffy
  • Francisco Noya
  • Wei Ming Chien

Detail Information

Publications25

  1. ncbi Simultaneous in situ detection of RNA, DNA, and protein using tyramide-coupled immunofluorescence
    Brian A Van Tine
    Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA
    Methods Mol Biol 292:215-30. 2005
    ..Finally, we describe the detection of nascent viral RNA transcripts simultaneously with integrated viral genomes or chromosomal domains in single cells or tissue sections...
  2. pmc Dynamic localization of the human papillomavirus type 11 origin binding protein E2 through mitosis while in association with the spindle apparatus
    Luan D Dao
    Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294 0005, USA
    J Virol 80:4792-800. 2006
    ..We conclude that this region of HPV E2C is critical for localization with the mitotic apparatus, enabling the HPV DNA to sustain persistent infections...
  3. pmc Conditionally activated E7 proteins of high-risk and low-risk human papillomaviruses induce S phase in postmitotic, differentiated human keratinocytes
    N Sanjib Banerjee
    Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294 0005, USA
    J Virol 80:6517-24. 2006
    ..These observations are entirely consistent with the fact that low-risk HPV genotypes replicate in the differentiated strata in patient specimens, as do the high-risk HPVs...
  4. pmc microRNAs are biomarkers of oncogenic human papillomavirus infections
    Xiaohong Wang
    Tumor Virus RNA Biology Section, Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892
    Proc Natl Acad Sci U S A 111:4262-7. 2014
    ..Further analyses indicate that an expression ratio ≥1.5 of miR-25/92a group over miR-22/29a group could serve as a cutoff value to distinguish normal cervix from CIN and from CIN to CC. ..
  5. pmc Phylogenetic considerations in designing a broadly protective multimeric L2 vaccine
    Subhashini Jagu
    Department of Pathology, Johns Hopkins University, Baltimore, Maryland, USA
    J Virol 87:6127-36. 2013
    ....
  6. pmc HPV-18 E6 mutants reveal p53 modulation of viral DNA amplification in organotypic cultures
    Eun Young Kho
    Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294 0005, USA
    Proc Natl Acad Sci U S A 110:7542-9. 2013
    ....
  7. pmc Ubp-M serine 552 phosphorylation by cyclin-dependent kinase 1 regulates cell cycle progression
    Yang Xu
    Department of Biochemistry and Molecular Genetics University of Alabama at Birmingham Birmingham, AL USA State Key Laboratory of Cellular and Molecular Biology College of Life Science Hebei Normal University Shijiazhuang, China
    Cell Cycle 12:3219-27. 2013
    ..Therefore, these studies confirm that Ubp-M is phosphorylated at S552 and identify CDK1 as the enzyme responsible for the phosphorylation. Importantly, this study specifically links Ubp-M S552P to cell cycle G 2/M phase progression...
  8. pmc Comprehensive control of human papillomavirus infections and related diseases
    F Xavier Bosch
    Cancer Epidemiology Research Program CERP, Institut Català d Oncologia Catalan Institute of Oncology ICO, IDIBELL, L Hospitalet de Llobregat Barcelona, Spain Electronic address
    Vaccine 31:I1-31. 2013
    ..Additional details on each subtopic and full information regarding the supporting literature references may be found in the original chapters. ..
  9. ncbi Comprehensive control of human papillomavirus infections and related diseases
    F Xavier Bosch
    Cancer Epidemiology Research Program CERP, Institut Català d Oncologia Catalan Institute of Oncology ICO, IDIBELL, L Hospitalet de Llobregat Barcelona, Spain Electronic address
    Vaccine 31:F1-31. 2013
    ..Additional details on each subtopic and full information regarding the supporting literature references may be found in the original chapters. ..
  10. ncbi Comprehensive control of human papillomavirus infections and related diseases
    F Xavier Bosch
    Cancer Epidemiology Research Program CERP, Institut Català d Oncologia Catalan Institute of Oncology ICO, IDIBELL, L Hospitalet de Llobregat Barcelona, Spain Electronic address
    Vaccine 31:G1-G31. 2013
    ..Additional details on each subtopic and full information regarding the supporting literature references may be found in the original chapters. ..
  11. ncbi Comprehensive control of human papillomavirus infections and related diseases
    F Xavier Bosch
    Cancer Epidemiology Research Program CERP, Institut Català d Oncologia Catalan Institute of Oncology ICO, IDIBELL, L Hospitalet de Llobregat Barcelona, Spain Electronic address
    Vaccine 31:H1-H31. 2013
    ..Additional details on each subtopic and full information regarding the supporting literature references may be found in the original chapters. ..
  12. pmc Adipose-derived stromal cells accelerate wound healing in an organotypic raft culture model
    Sherry S Collawn
    Division of Plastic Surgery, University of Alabama, Birmingham, AL, USA
    Ann Plast Surg 68:501-4. 2012
    ..In conclusion, these data provide support to our hypothesis that ADSCs are an excellent and readily available source of factors necessary for accelerated wound healing and tissue regeneration...
  13. pmc Construction of a full transcription map of human papillomavirus type 18 during productive viral infection
    Xiaohong Wang
    Tumor Virus RNA Biology Section, HIV and AIDS Malignancy Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
    J Virol 85:8080-92. 2011
    ..Collectively, a full HPV18 transcription map constructed from this report will lead us to further understand HPV18 gene expression and virus oncogenesis...
  14. pmc PIK3CA-mediated PI3-kinase signalling is essential for HPV-induced transformation in vitro
    Florianne E Henken
    Department of Pathology, Unit of Molecular Pathology, VU University Medical Center, PO Box 7057, 1007 MB Amsterdam, The Netherlands
    Mol Cancer 10:71. 2011
    ..In this study we experimentally assessed the role of the PI3-kinase pathway and its regulator PIK3CA, which is frequently altered in cervical cancer, in HPV-induced transformation...
  15. pmc Nonconserved lysine residues attenuate the biological function of the low-risk human papillomavirus E7 protein
    Nicholas J Genovese
    University of Alabama at Birmingham, Biochemistry and Molecular Genetics, Birmingham, AL 35294 0005, USA
    J Virol 85:5546-54. 2011
    ..Thus, these nonconserved residues are primarily responsible for the differential abilities of LR and HR HPV E7 proteins to promote unscheduled DNA replication in organotypic raft cultures...
  16. pmc Human papillomavirus (HPV) E7 induces prolonged G2 following S phase reentry in differentiated human keratinocytes
    N Sanjib Banerjee
    Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, Alabama 35294 0005, USA
    J Biol Chem 286:15473-82. 2011
    ..Collectively, this study has revealed the mechanisms by which E7 induces prolonged G(2) phase in the differentiated cells following S phase induction...
  17. ncbi The natural history of human papillomavirus infections of the mucosal epithelia
    Louise T Chow
    Department of Biochemistry and Molecular Genetics, The University of Alabama at Birmingham, 35294 0005, USA
    APMIS 118:422-49. 2010
    ..Finally, we provide our thoughts about treatments for HPV-induced diseases, present and future...
  18. ncbi Anal-rectal cytology: correlation with human papillomavirus status and biopsy diagnoses in a population of HIV-positive patients
    Sarah M Bean
    Duke University Medical Center, Durham, NC 27710, USA
    J Low Genit Tract Dis 14:90-6. 2010
    ....
  19. pmc Mitogen-activated protein kinases activate the nuclear localization sequence of human papillomavirus type 11 E1 DNA helicase to promote efficient nuclear import
    Jei Hwa Yu
    Department of Biochemistry and Molecular Genetics, McCallum Building, University of Alabama at Birmingham, 1918 University Boulevard, Birmingham, Alabama 35294 0005, USA
    J Virol 81:5066-78. 2007
    ..In contrast, E1 proteins mutated in the MAPK docking motifs were completely inactive in transient replication, an indication that additional properties were adversely affected by those changes...
  20. pmc Remodeling of the human papillomavirus type 11 replication origin into discrete nucleoprotein particles and looped structures by the E2 protein
    Jeonggu Sim
    Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599 7295, USA
    J Mol Biol 375:1165-77. 2008
    ..The role of these structures in the modeling and regulation of the HPV-11 ori is discussed...
  21. pmc Casein kinase II motif-dependent phosphorylation of human papillomavirus E7 protein promotes p130 degradation and S-phase induction in differentiated human keratinocytes
    Nicholas J Genovese
    University of Alabama at Birmingham, Biochemistry and Molecular Genetics, 1918 University Boulevard, McCallum Building 510, Birmingham, AL 35294 0005, USA
    J Virol 82:4862-73. 2008
    ..Collectively, these data support the notion that p130 controls the homeostasis of the differentiated keratinocytes and is therefore targeted by E7 for degradation to establish conditions permissive for viral DNA amplification...
  22. pmc Robust production and passaging of infectious HPV in squamous epithelium of primary human keratinocytes
    Hsu Kun Wang
    Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA
    Genes Dev 23:181-94. 2009
    ..This system will be invaluable for HPV genetic dissection and serves as a faithful ex vivo model for investigating infections and interventions...
  23. pmc Oncogenic HPV infection interrupts the expression of tumor-suppressive miR-34a through viral oncoprotein E6
    Xiaohong Wang
    HIV and AIDS Malignancy Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA
    RNA 15:637-47. 2009
    ..Together, this is the first time a viral oncoprotein has been shown to regulate cellular miRNA expression. Our data have provided new insights into mechanisms by which high-risk HPVs contribute to the development of cervical cancer...
  24. ncbi A highly efficient system to produce infectious human papillomavirus: Elucidation of natural virus-host interactions
    Louise T Chow
    Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294, USA
    Cell Cycle 8:1319-23. 2009
    ..We provide additional insights into the viral life cycle and contrast them to conclusions derived from experiments in cell lines...