AAV-2 Rep78 and HPV-16 E1 interact in vitro, modulating their ATPase activity.
Adeno-associated virus (AAV) is a nonpathogenic single-stranded human parvovirus which usually requires the presence of a "helper" virus for strong DNA replication. In addition to adeno- and herpes viruses, human papillomavirus (HPV) can serve as an AAV helper. We recently published that HPV type 16 (HPV-16) E1 protein contributes significantly as an individual helper gene for AAV-2 DNA replication and transcription. As Rep78 and E1 are the corresponding DNA helicase/replication proteins of AAV and HPV, respectively, and Rep78 and E1 have a degree of homology, we assayed whether these two proteins interact physically. The full length proteins were purified from bacteria as GST-E1 and MBP-Rep78 and used in five assays to observe Rep78-E1 interactions. All five assays (pull-down, coimmunoprecipitation, enzyme-linked immunosorbent assay (ELISA), chemical cross-linking, and ATPase activity) provided evidence consistent with Rep78-E1 interaction. Most intriguing, an overall decrease in ATPase activity was observed when both proteins were present together. These data strongly suggest that E1 and Rep78 interact and that this interaction modulates at least some of their individual biochemical functions. This study adds to our understanding of AAV-HPV interaction biology, E1's modulation of Rep78 biochemistry, Rep78's modulation of E1 biochemistry and provides initial clues which may lead to the underlying mechanism of HPV E1 helper function for AAV DNA replication.
Bandyopadhyay
Sarmistha
S
Department of Internal Medicine, Gene Therapy Program, University of Arkansas for Medical Sciences, 4301 West Markham Street, Little Rock, Arkansas 72205, USA.
Raney
Kevin D
KD
Liu
Yong
Y
Hermonat
Paul L
PL
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D047468:Immunoprecipitation; D009856:Oncogene Proteins, Viral / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism*
D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; C093478:E1 protein, Human papillomavirus 16; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C068734:rep proteins, Adeno-associated virus 2; D000251:Adenosine Triphosphatases