Two distinct regions of the BPV1 E1 replication protein interact with the activation domain of E2.
Papillomavirus E1 and E2 proteins co-operation in viral DNA replication is mediated by protein-protein interactions that lead to formation of an E1-E2 complex. To identify the domains involved, portions of the two proteins were expressed as fusions to the DNA-binding protein LexA or the transactivation domain of VP16 and analyzed by the yeast two-hybrid system. The C-terminal 266 amino acids of BPV1 E1 (E1C266) interacted strongly with E2 in the yeast system and in a mammalian two-hybrid assay. VP16-E1C266 interacted with a region encompassing amino acids 1-200 of the transactivation domain of E2 that was fused to LexA. The interaction between E1 full length and E2 was clearly observed only when E1 was expressed as LexA-E1 chimera. In addition, we found that in the LexA context also the N-terminal region encompassing the first 340 amino acids of E1 (E1N340) interacted with E2 full length. The interactions of E1N340 and E1C266 with E2 were confirmed also by in vitro binding studies. These observations demonstrate that two distinct regions of E1 mediate the interaction with E2 in vivo.
Moscufo
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N
Department of Dermatology, New England Medical Center and Tufts University School of Medicine, Boston, MA, USA. nmoscufo@iss.it
Sverdrup
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F
Breiding
D E
DE
Androphy
E J
EJ
D000818:Animals; D052688:Bovine papillomavirus 1 / Q000737:chemistry / Q000502:physiology; D002417:Cattle; D004265:DNA Helicases / Q000737:chemistry; D004279:DNA, Viral / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry*; D004742:Enhancer Elements, Genetic / Q000502:physiology; D015534:Trans-Activators / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D014764:Viral Proteins / Q000737:chemistry*; D014779:Virus Replication / Q000502:physiology
D004279:DNA, Viral; D004268:DNA-Binding Proteins; C069814:E1 protein, Bovine papillomavirus; C051550:E2 protein, Bovine papillomavirus; D015534:Trans-Activators; D014764:Viral Proteins; D004265:DNA Helicases