HPV11 E6 mutation by overexpression of APOBEC3A and effects of interferon-ω on APOBEC3s and HPV11 E6 expression in HPV11.HaCaT cells

Virol J. 2017 Nov 3;14(1):211. doi: 10.1186/s12985-017-0878-2.

Abstract

Background: Condyloma acuminatum, infected by low-risk human papillomaviruses (e.g., HPV6 and HPV11), is one of the most widespread sexually transmitted diseases. Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 proteins (APOBEC3s, A3s) are cellular cytidine deaminases acting as antiviral factors through hypermutation of viral genome. However, it remains unknown whether A3s results in HPV11 gene mutations and interferon-ω (IFN-ω) exhibits antiviral activities through the A3s system. Here we investigated whether enhanced APOBEC3A (A3A) resulted in the E6 gene mutations and explore the effects of recombinant human interferon-ω (rhIFN-ω) on A3s/E6 expression in HaCaT keratinocytes containing the genome of HPV 11 (HPV11.HaCaT cells).

Methods: A3A-overexpressed HPV11.HaCaT (A3A-HPV11.HaCaT) cells were established by lentiviral infection and verified by immunofluorescence and western-blotting. Cell cycle, E6 gene mutations, APOBEC3s/E6 gene expression and subcellular localization were detected by FACS, 3D-PCR and sequencing, qRT-PCR and immunofluorescence respectively.

Results: The results suggested that A3A-HPV11.HaCaT cells were successfully established. Enhanced A3A induced S-phase arrest, G > A/C > T mutations and obvious reduction of E6 mRNA expression. A3A/A3B mRNA expression was up-regulated at 6 h and 12 h and obvious A3A staining existed throughout HPV11.HaCaT cells after rhIFN-ω treatment. RhIFN-ω could also inhibit mRNA expression of HPV11 E6 significantly.

Conclusions: Enhanced A3A repressed HPV11 E6 expression through gene hypermutation, and rhIFN-ω might be an effective agent against HPV11 infection by up-regulation of A3A.

Keywords: Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3; Condylomata acuminata; Early gene 6; HaCaT keratinocytes; Human papillomavirus 11; Interferon-ω.

MeSH terms

  • Cell Cycle
  • Cell Line
  • Condylomata Acuminata / immunology*
  • Condylomata Acuminata / virology
  • Cytidine Deaminase / genetics
  • Cytidine Deaminase / metabolism*
  • Gene Expression
  • Human papillomavirus 11 / genetics*
  • Humans
  • Interferon Type I / metabolism*
  • Keratinocytes / virology
  • Mutation
  • Oncogene Proteins, Viral / genetics*
  • Proteins / genetics
  • Proteins / metabolism*
  • RNA, Messenger
  • Up-Regulation

Substances

  • E6 protein, Human papillomavirus type 11
  • Interferon Type I
  • Oncogene Proteins, Viral
  • Proteins
  • RNA, Messenger
  • interferon omega 1
  • APOBEC3A protein, human
  • Cytidine Deaminase