Inhibition of human papillomavirus DNA replication by small molecule antagonists of the E1-E2 protein interaction.
Human papillomavirus (HPV) DNA replication is initiated by recruitment of the E1 helicase by the E2 protein to the viral origin. Screening of our corporate compound collection with an assay measuring the cooperative binding of E1 and E2 to the origin identified a class of small molecule inhibitors of the protein interaction between E1 and E2. Isothermal titration calorimetry and changes in protein fluorescence showed that the inhibitors bind to the transactivation domain of E2, the region that interacts with E1. These compounds inhibit E2 of the low risk HPV types 6 and 11 but not those of high risk HPV types or of cottontail rabbit papillomavirus. Functional evidence that the transactivation domain is the target of inhibition was obtained by swapping this domain between a sensitive (HPV11) and a resistant (cottontail rabbit papillomavirus) E2 type and by identifying an amino acid substitution, E100A, that increases inhibition by approximately 10-fold. This class of inhibitors was found to antagonize specifically the E1-E2 interaction in vivo and to inhibit HPV DNA replication in transiently transfected cells. These results highlight the potential of the E1-E2 interaction as a small molecule antiviral target.
Department of Biological Sciences and the Department of Chemistry, Boehringer Ingelheim Ltd., Laval H7S 2G5, Canada.
Peter W
PW
White
Steve
S
Titolo
Karine
K
Brault
Louise
L
Thauvette
Alex
A
Pelletier
Ewald
E
Welchner
Lise
L
Bourgon
Louise
L
Doyon
William W
WW
Ogilvie
Christiane
C
Yoakim
Michael G
MG
Cordingley
Jacques
J
Archambault
D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D016466:CHO Cells; D012777:Cottontail rabbit papillomavirus / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D006224:Cricetinae; D004261:DNA Replication / Q000187:drug effects*; D004279:DNA, Viral / Q000096:biosynthesis* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D027383:Papillomaviridae / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D011817:Rabbits; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation; D014162:Transfection; D014764:Viral Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*
D004279:DNA, Viral; D004268:DNA-Binding Proteins; C084304:E1 protein, Human papillomavirus type 11; C118861:E1 protein, Human papillomavirus type 6; C117355:E2 protein, Human papillomavirus type 11; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins