Human papillomaviruses (HPV) form a large family of viruses some of which are associated with various forms of cancer, including cervical carcinoma. They induce more than seven per cent of all cancers in women worldwide. Especially, HPV pose a burden to women in underdeveloped regions in the world due to a lack of access to cancer screening programs. The long term goal of this research is to understand early events of HPV infection with the focus on events occurring on the cell surface. The major capsid protein, L1, of HPV type 16 is primarily responsible for initial binding of virus to heparan sulfate proteoglycan (HSPG) present on cell surface and extracellular matrix (ECM). Attachment to HSPG induces a shift in the conformation of L1 and the minor capsid protein, L2, both of which we can now detect with monoclonal antibodies. X-ray structure analysis in collaboration with X. Chen identified several HSPG binding sites on the viral capsid. Our preliminary data suggest their sequential use in primary attachment and secondary binding events as well for inducing conformational shifts. In addition, we collected evidence for an involvement of Cyclophilin B (CyPB) in two distinct steps during HPV infection: mediating L2 conformational changes on the cell surface and facilitating uncoating of the viral genome.
Aim 1 will test our hypothesis that two HS binding sites on the viral capsid are sequentially used during cell surface events of HPV infection. This will be achieved by a structure-guided mutagenesis of HS binding sites.
Aim 2 is intended to delineate the function of Cyclophilins in HPV infection using siRNA knock down and small molecule inhibitors combined with biochemical and cell biological approaches.
Aim 3 is dedicated to understand the processes underlying the conformational shifts affecting both capsid proteins in molecular detail. Understanding these events in detail will allow the development of interfering strategies to fight HPV-induced malignant lesions.

Public Health Relevance

HPV induce a variety of cancers, including cervical carcinoma. We will investigate how the virus exploits host cell factors for attachment to host cells and subsequent events leading to successful infection. This should allow identification of cellular targets for drug therapies to prevent HPV induced cancers.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI081809-05
Application #
8500124
Study Section
Virology - A Study Section (VIRA)
Program Officer
Park, Eun-Chung
Project Start
2009-07-17
Project End
2014-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
5
Fiscal Year
2013
Total Cost
$337,425
Indirect Cost
$107,101
Name
Louisiana State University Hsc Shreveport
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
095439774
City
Shreveport
State
LA
Country
United States
Zip Code
71103
DiGiuseppe, Stephen; Bienkowska-Haba, Malgorzata; Hilbig, Lydia et al. (2014) The nuclear retention signal of HPV16 L2 protein is essential for incoming viral genome to transverse the trans-Golgi network. Virology 458-459:93-105
Richards, Kathleen F; Bienkowska-Haba, Malgorzata; Dasgupta, Jhimli et al. (2013) Multiple heparan sulfate binding site engagements are required for the infectious entry of human papillomavirus type 16. J Virol 87:11426-37
Spoden, Gilles; Kuhling, Lena; Cordes, Nicole et al. (2013) Human papillomavirus types 16, 18, and 31 share similar endocytic requirements for entry. J Virol 87:7765-73
Sapp, Martin J (2013) HPV virions hitchhike a ride on retromer complexes. Proc Natl Acad Sci U S A 110:7116-7
Dasgupta, Jhimli; Bienkowska-Haba, Malgorzata; Ortega, Marcos E et al. (2011) Structural basis of oligosaccharide receptor recognition by human papillomavirus. J Biol Chem 286:2617-24
Bienkowska-Haba, Malgorzata; Sapp, Martin (2011) The cytoskeleton in papillomavirus infection. Viruses 3:260-71
Sapp, Martin; Bienkowska-Haba, Malgorzata (2009) Viral entry mechanisms: human papillomavirus and a long journey from extracellular matrix to the nucleus. FEBS J 276:7206-16