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DNA replicative functions of highly-expressed, codon-optimized human papillomavirus proteins E1 and E2
Xin-Min Wang1, Kathrin U Jansen, William L McClements
1Department of Microbial Vaccine Research, P O Box 4, West Point, PA 19486, USA.
Journal of Virological Methods
|February 5, 2003
Summary
Researchers assessed human papillomavirus (HPV) E1 and E2 mutant proteins for vaccine potential. An HPV 16 E1 mutant failed to support DNA replication, while an E2 mutant showed only a two-fold reduction in activity.
Area of Science:
- Virology
- Molecular Biology
- Vaccine Development
Background:
- Human papillomavirus (HPV) DNA replication is essential and relies on the E1 and E2 early proteins.
- Mutant HPV proteins are being explored as potential vaccine candidates.
Purpose of the Study:
- To evaluate the biological activity and DNA replicative functions of HPV 16 E1 and E2 mutant proteins.
- To determine the suitability of these mutants for vaccine development.
Main Methods:
- Development of a sensitive real-time PCR assay to monitor HPV origin-of-replication-driven DNA synthesis.
- Transient transfection assays to assess the DNA replicative functions of wild-type and mutant HPV 16 E1 and E2 proteins.
Main Results:
- The HPV 16 E1 mutant (W439R, G482D) did not support HPV origin-driven DNA synthesis under the assay conditions.
- An HPV 16 E2 mutant (E39A) showed only a two-fold reduction in DNA replication activity, contrary to previous reports.
Conclusions:
- The HPV 16 E1 mutant is significantly impaired in its DNA replication function.
- The HPV 16 E2 mutant (E39A) is not sufficiently inactivated for use in vaccines relying on endogenous protein expression due to its residual replication activity.