Effect of bovine papillomavirus E2 protein-specific monoclonal antibodies on papillomavirus DNA replication.
The bovine papillomavirus type 1 (BPV-1) E2 protein is the master regulator of papillomavirus replication and transcription. We have raised a panel of monoclonal antibodies (MAbs) against the BPV-1 E2 protein and used them to probe the structure and function of the protein. Five MAbs reacted with linear epitopes, and four MAbs recognized conformation-dependent epitopes which mapped within the C-terminal DNA-binding and dimerization domain. MAb 1E2 was able to recognize the replication- and transactivation-defective but not the competent conformation of the transactivation domain of the E2 protein. MAb 5H4 prevented efficiently the formation of E2-DNA as well as E2-dependent E1-E2-origin complexes and also dissociated preformed complexes in a concentration-dependent manner. Cotransfection of several MAbs with the BPV-1 minimal origin plasmid pUCAlu into CHO4.15 cells resulted in a dose-dependent inhibition of replication. Inhibition of replication by MAb 5H4 and the Fab' fragment of 5H4 correlated with their ability to dissociate the E2 protein from the DNA. MAb 3F12 and MAbs 1H10 and 1E4, directed against the hinge region, were also capable of inhibiting BPV-1 origin replication in CHO4.15 cells. However, the Fab' fragments of 1H10 and 3F12 had no effect in the transient replication assay. These data suggest that MAbs directed against the hinge region sterically hinder the inter- or intramolecular interactions required for the replication activity of the E2 protein.
Kurg
R
R
Department of Microbiology and Virology, Institute of Molecular and Cell Biology, Tartu, Estonia.
Parik
J
J
Juronen
E
E
Sedman
T
T
Abroi
A
A
Liiv
I
I
Langel
U
U
Ustav
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D052688:Bovine papillomavirus 1 / Q000502:physiology*; D016466:CHO Cells; D019556:COS Cells; D006224:Cricetinae; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000276:immunology / Q000502:physiology*; D018604:Epitope Mapping; D005260:Female; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D014764:Viral Proteins / Q000276:immunology / Q000502:physiology*; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D004268:DNA-Binding Proteins; C051550:E2 protein, Bovine papillomavirus; D007140:Immunoglobulin Fab Fragments; D014764:Viral Proteins