Amino-terminal domains of the bovine papillomavirus type 1 E1 and E2 proteins participate in complex formation.
Interaction between the E1 and E2 papillomavirus proteins appear to play an important role in viral DNA replication, although the exact domains of each protein involved in this interaction have not been identified. Using bovine papillomavirus type 1 (BPV-1) as a model for examining interactions between E1 and E2, we have used the two-hybrid and glutathione S-transferase (GST) fusion systems to map domains of BPV-1 E1 and E2 that interact in vivo and in vitro. In the two-hybrid system experiments, portions of BPV-1 E2 were expressed in Saccharomyces cerevisiae as LexA fusion proteins, which were tested for interaction with various domains of BPV-1 E1. These assays indicated that domains sufficient for E1-E2 interaction are present within the amino-terminal 250 amino acids of E1 and within the first 91 amino acids of E2. Interestingly, a LexA fusion protein that included amino acid residues 53 to 161 of BPV E2 demonstrated transcriptional activation in this system. In vitro binding assays using combinations of BPV-1 E1-GST fusion proteins and BPV-1 E2 expressed by in vitro translation confirmed the observations from the yeast system; a GST fusion protein containing the first 222 amino acids of BPV-1 E1 bound specifically to full-length BPV-1 E2 in vitro. Furthermore, E1(1-222)-GST bound to forms of E2 deleted of the carboxy-terminal DNA binding-dimerization domain, suggesting that E2 dimerization is not required for this interaction. Finally, in vitro interaction between E1-GST and E2 was observed at 22 degrees C but not at 4 degrees C.
Benson
J D
JD
Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Howley
P M
PM
D001426:Bacterial Proteins / Q000737:chemistry; D052688:Bovine papillomavirus 1 / Q000737:chemistry*; D003080:Cold Temperature; D004261:DNA Replication; D010446:Peptide Fragments / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012697:Serine Endopeptidases; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000737:chemistry* / Q000502:physiology; D014779:Virus Replication; D000099160:LexA Repressor Protein
D001426:Bacterial Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D012697:Serine Endopeptidases; D014764:Viral Proteins; D000099160:LexA Repressor Protein