title: Activation of NF-κB by human papillomavirus 16 E1 limits E1-dependent viral replication through degradation of E1.
abstract: NF-κB is a family of transcription factors that regulate gene expression involved in many processes, such as the inflammatory response and cancer progression. Little is known about associations of NF-κB with the human papillomavirus (HPV) life cycle. We have developed a tissue culture system to conditionally induce E1-dependent replication of the human papillomavirus 16 (HPV16) genome in human cervical keratinocytes and found that expression of HPV16 E1, a viral helicase, results in reduction of IκBα and subsequent activation of NF-κB in a manner dependent on helicase activity. Exogenous expression of a degradation-resistant mutant of IκBα, which inhibits the activation of NF-κB, enhanced E1-dependent replication of the viral genome. Wortmannin, a broad inhibitor of phosphoinositide 3-kinases (PI3Ks), and, to a lesser extent, VE-822, an ATR kinase inhibitor, but not KU55933, an ATM kinase inhibitor, suppressed the activation of NF-κB and augmented E1-dependent replication of the HPV16 genome. Interestingly, the enhancement of E1-dependent replication of the viral genome was associated with increased stability of E1 in the presence of wortmannin as well as the IκBα mutant. Collectively, we propose that expression of E1 induces NF-κB activation at least in part through the ATR-dependent DNA damage response and that NF-κB in turn limits E1-dependent replication of HPV16 through degradation of E1, so that E1 and NF-κB may constitute a negative feedback loop.
  
  
  OBJECTIVE
  A major risk factor in human papillomavirus (HPV)-associated cancers is persistent infection with high-risk HPVs. To eradicate viruses from infected tissue, it is important to understand molecular mechanisms underlying the establishment and maintenance of persistent infection. In this study, we obtained evidence that human papillomavirus 16 (HPV16) E1, a viral DNA helicase essential for amplification of the viral genomes, induces NF-κB activation and that this limits E1-dependent genome replication of HPV16. These results suggest that NF-κB mediates a negative feedback loop to regulate HPV replication and that this feedback loop could be associated with control of the viral copy numbers. We could thus show for the first time that NF-κB activity is involved in the establishment and maintenance of persistent HPV infection.
authors.affiliation: Division of Carcinogenesis and Cancer Prevention, National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Nakahara
authors.affiliation: Division of Carcinogenesis and Cancer Prevention, National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Tanaka
authors.affiliation: Division of Carcinogenesis and Cancer Prevention, National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Ohno
authors.affiliation: Division of Carcinogenesis and Cancer Prevention, National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: Nagayasu
authors.initials: N
authors.lastname: Egawa
authors.affiliation: Division of Carcinogenesis and Cancer Prevention, National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yugawa
authors.affiliation: Division of Carcinogenesis and Cancer Prevention, National Cancer Center Research Institute, Tokyo, Japan tkiyono@ncc.go.jp.
authors.forename: Tohru
authors.initials: T
authors.lastname: Kiyono
mesh_terms: D002478:Cells, Cultured; D054884:Host-Pathogen Interactions; D052162:Human papillomavirus 16 / Q000276:immunology* / Q000502:physiology; D006801:Humans; D015603:Keratinocytes / Q000821:virology; D016328:NF-kappa B / Q000276:immunology* / Q000378:metabolism*; D009856:Oncogene Proteins, Viral / Q000276:immunology* / Q000378:metabolism*; D059748:Proteolysis; D014779:Virus Replication
chemical_list: C093478:E1 protein, Human papillomavirus 16; D016328:NF-kappa B; D009856:Oncogene Proteins, Viral