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Journal of Virology, September 2007, p. 9922-9931, Vol. 81, No. 18
0022-538X/07/$08.00+0     doi:10.1128/JVI.00988-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Human Papillomavirus Type 31 Uses a Caveolin 1- and Dynamin 2-Mediated Entry Pathway for Infection of Human Keratinocytes{triangledown}

Jessica L. Smith, Samuel K. Campos, and Michelle A. Ozbun*

Department of Molecular Genetics and Microbiology, The University of New Mexico School of Medicine, Albuquerque, New Mexico 87131

Received 7 May 2007/ Accepted 29 June 2007

Papillomaviruses are species-specific and epitheliotropic DNA viruses that cause tumors in their natural hosts. Certain infections with genital human papillomavirus (HPV) types are causally related to cervical cancer development. Most papillomaviruses are thought to infect cells via a clathrin-dependent pathway, yet no studies have determined the entry route in permissive host epithelial cells. Employing fluorescently labeled and native virions, we tested the effects of dominant-negative and biochemical inhibitors of cellular endocytosis pathways. Infections of human keratinocytes, a natural host cell type for HPVs, were assessed visually and by infectious entry assays. We found that HPV type 31 (HPV31) entry and initiation of early infection events require both caveolin 1 and dynamin 2 and occur independently of clathrin-mediated endocytosis. Treatment with chlorpromazine and filipin had opposing effects on HPV31 and HPV16 infection. HPV31 entry was remarkably slow, with a half-time of {approx}14 h, whereas the entry half-time of HPV16 was 4 h. Consistent with a caveola-mediated entry pathway for HPV31, the virions associated with detergent-resistant lipid rafts. During a 16-h microscopic tracking of HPV31 and HPV16 virions, no colocalization of the two viral types was observed. These data suggest that HPV31 and HPV16 virions use distinct routes for host epithelial cell entry.

* Corresponding author. Mailing address: Department of Molecular Genetics and Microbiology, The University of New Mexico School of Medicine, Albuquerque, NM 87131. Phone: (505) 272-4950. Fax: (505) 272-9912. E-mail: mozbun{at}salud.unm.edu

{triangledown} Published ahead of print on 11 July 2007.

Journal of Virology, September 2007, p. 9922-9931, Vol. 81, No. 18
0022-538X/07/$08.00+0     doi:10.1128/JVI.00988-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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