This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ozbun, M. A. Search for Related Content PubMed PubMed Citation Articles by Ozbun, M. A.

 Previous Article  |  Next Article 

Journal of Virology, November 2002, p. 11291-11300, Vol. 76, No. 22
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.22.11291-11300.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Human Papillomavirus Type 31b Infection of Human Keratinocytes and the Onset of Early Transcription

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

Received 22 May 2002/ Accepted 12 August 2002

Human papillomaviruses (HPVs) cause a number of human tumors and malignancies, including cervical cancers. Epithelial differentiation is required for the complete HPV life cycle and can be achieved using the organotypic (raft) culture system. The CIN-612 9E cell line maintains episomal copies of HPV type 31b (HPV31b), an HPV type associated with cervical cancers. When grown in the raft system, CIN-612 9E cells form a differentiated epithelium such that infectious virions can be synthesized. Many aspects of the later stages of the HPV31b life cycle have been investigated in CIN-612 9E raft tissues. We used a biologically contained homogenization system for efficient virion extraction from raft epithelial tissues. Purified HPV31b virions were used to infect low-passage-number human foreskin keratinocytes and a variety of epithelial cell lines. Newly synthesized, spliced HPV31b transcripts were detected by reverse transcription and PCR (RT-PCR) following HPV31b infection. HPV31b infection was most efficient and reproducible in HaCaT cells. The onset of viral transcription following infection was also investigated using RT-PCR techniques. Spliced E1_*I,E2 RNAs were present as early as 4 h postinfection (p.i.), whereas the other major viral transcripts were detected by 8 to 10 h p.i. Furthermore, we characterized the structures and temporal expression of seven novel spliced early transcripts expressed following infection.

This article has been cited by other articles:

• Smith, J. L., Campos, S. K., Ozbun, M. A. (2007). Human Papillomavirus Type 31 Uses a Caveolin 1- and Dynamin 2-Mediated Entry Pathway for Infection of Human Keratinocytes. J. Virol. 81: 9922-9931 [Abstract] [Full Text]  
• Carson, A., Khan, S. A. (2006). Characterization of Transcription Factor Binding to Human Papillomavirus Type 16 DNA during Cellular Differentiation. J. Virol. 80: 4356-4362 [Abstract] [Full Text]  
• Pyeon, D., Lambert, P. F., Ahlquist, P. (2005). Production of infectious human papillomavirus independently of viral replication and epithelial cell differentiation. Proc. Natl. Acad. Sci. USA 102: 9311-9316 [Abstract] [Full Text]  
• Patterson, N. A., Smith, J. L., Ozbun, M. A. (2005). Human Papillomavirus Type 31b Infection of Human Keratinocytes Does Not Require Heparan Sulfate. J. Virol. 79: 6838-6847 [Abstract] [Full Text]  
• Holmgren, S. C., Patterson, N. A., Ozbun, M. A., Lambert, P. F. (2005). The Minor Capsid Protein L2 Contributes to Two Steps in the Human Papillomavirus Type 31 Life Cycle. J. Virol. 79: 3938-3948 [Abstract] [Full Text]  
• Lee, J. H., Yi, S. M. P., Anderson, M. E., Berger, K. L., Welsh, M. J., Klingelhutz, A. J., Ozbun, M. A. (2004). Propagation of infectious human papillomavirus type 16 by using an adenovirus and Cre/LoxP mechanism. Proc. Natl. Acad. Sci. USA 101: 2094-2099 [Abstract] [Full Text]  
• Kim, K., Garner-Hamrick, P. A., Fisher, C., Lee, D., Lambert, P. F. (2003). Methylation Patterns of Papillomavirus DNA, Its Influence on E2 Function, and Implications in Viral Infection. J. Virol. 77: 12450-12459 [Abstract] [Full Text]  
• Oberg, D., Collier, B., Zhao, X., Schwartz, S. (2003). Mutational Inactivation of Two Distinct Negative RNA Elements in the Human Papillomavirus Type 16 L2 Coding Region Induces Production of High Levels of L2 in Human Cells. J. Virol. 77: 11674-11684 [Abstract] [Full Text]