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Tracking Replicating HPV Genomes in Proliferating Keratinocytes.

Jonathan R Shin1, Ivan Avilov2, Mario Schelhaas2

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This summary is machine-generated.

Researchers developed a new method to track human papillomavirus (HPV) DNA in living cells using ANCHOR technology. This allows real-time monitoring of HPV18 genome replication and distribution in keratinocytes.

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Area of Science:

  • Virology
  • Molecular Biology
  • Cell Biology

Background:

  • Human papillomavirus (HPV) genomes function as minichromosomes during persistent infections.
  • Monitoring viral genome dynamics in living cells is challenging due to the genome's small size and compact structure.

Purpose of the Study:

  • To develop a novel method for detecting and tracking HPV18 genomes in living cells using ANCHOR technology.
  • To visualize the replication, maintenance, and partitioning of HPV18 genomes in proliferating keratinocytes.

Main Methods:

  • Incorporation of the ANCHOR cis element into the HPV18 genome.
  • Expression of ParB-GFP protein from an HPV18-dependent replicon containing the viral origin and enhancer.
  • Utilizing a neomycin resistance gene for selection in both bacterial and eukaryotic cells.
  • Real-time tracking of fluorescently labeled HPV18 genomes in live keratinocytes.

Main Results:

  • The ANCHOR technology enabled indirect monitoring of HPV18 genomes via fluorescent protein detection.
  • Replicating HPV18-ANCH3 genomes were tracked in real-time within proliferating keratinocytes.
  • HPV18 genomes were observed to partition relatively equally to daughter cells during cell division through random attachment to host chromosomes.

Conclusions:

  • The developed molecular tool facilitates the study of HPV genome dynamics in live cells.
  • This method provides insights into HPV genome maintenance, tethering, and amplification during infection.
  • The findings elucidate the partitioning mechanisms of HPV18 DNA in dividing host cells.