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Transforming activity of E5a protein of human papillomavirus type 6 in NIH 3T3 and C127 cells
Abstract
Human papillomavirus type 6 (HPV-6) is the etiologic agent of genital warts and recurrent respiratory papillomatosis. We are investigating the mechanism by which this virus stimulates cell proliferation during infection. In this paper, we report that the E5a gene of HPV-6c, an independent isolate of HPV-11, is capable of transforming NIH 3T3 cells. The E5a open reading frame (ORF) was expressed under the control of the mouse metallothionein promoter in the expression vector pMt.neo.1, which also contains the gene for G418 resistance. Transfected cells were selected for G418 resistance and analyzed for a transformed phenotype. The transformed NIH 3T3 cells overgrew a confluent monolayer, had an accelerated generation time, and were anchorage independent. In contrast, E5a did not induce foci in C127 cells, but C127 cells expressing E5a did form small colonies in suspension. The presence of the 12-kilodalton E5a gene product in the transformed NIH 3T3 cells was shown by immunoprecipitation and was localized predominantly to nuclei by an immunoperoxidase assay. A mutation in the E5a ORF was engineered to terminate translation. This mutant was defective for transformation, demonstrating that translation of the E5a ORF is required for biological activity. This is the first demonstration of a transforming oncogene in HPV-6, and the differential activity of E5a in these two cell lines should facilitate future investigations on the mechanism of transformation.
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“Initial studies with the expression of HPV-6 E5 in murine fibroblasts showed that it could induce the formation of colonies in soft agar [100]”
From The Not-So-Good, the Bad and the Ugly: HPV E5, E6 and E7 Oncoproteins in the Orchestration of Carcinogenesis (2021) · cites this paper as [100]
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Cited by
Basukala, O., & Banks, L. (2021) · Viruses · doi:10.3390/v13101892 · Cited by 137
Metadata
- Authors
- Chen, S. L., & Mounts, P.
- Affiliations
- Department of Immunology and Infectious Diseases, School of Hygiene and Public Health, John Hopkins University, Baltimore, Maryland 21205.
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 1990-07
- Volume
- 64
- Issue
- 7
- Pages
- 3226-3233
- ISSN
- 0022-538X
- 1098-5514
- Cited by
- 64
- References
- 1
- Language
- en
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