Beta human papillomavirus 8E6 promotes alternative end-joining

  1. Changkun Hu
  2. Taylor Bugbee
  3. Rachel Palinski
  4. Ibukun A Akinyemi
  5. Michael T McIntosh
  6. Thomas MacCarthy
  7. Sumita Bhaduri-McIntosh
  8. Nicholas Wallace  Is a corresponding author
  1. Fred Hutchinson Cancer Research Center, United States
  2. Kansas State University, United States
  3. University of Florida, United States
  4. Stony Brook University, United States

Abstract

Double strand breaks (DSBs) are one of the most lethal DNA lesions in cells. The E6 protein of beta-human papillomavirus (HPV8 E6) impairs two critical DSB repair pathways; homologous recombination (HR) and non-homologous end-joining (NHEJ). However, HPV8 E6 only delays DSB repair. How DSBs are repaired in cells with HPV8 E6 remains to be studied. We hypothesize that HPV8 E6 promotes a less commonly used DSB repair pathway, alternative end-joining (Alt-EJ). Using CAS9 based Alt-EJ reporters, we show that HPV8 E6 promotes Alt-EJ. Further, using small molecule inhibitors, CRISPR/CAS9 gene knockout, and HPV8 E6 mutant, we find that HPV8 E6 promotes Alt-EJ by binding p300, an acetyltransferase that facilitates DSB repair by HR and NHEJ. At least some of this repair occurs through a subset of Alt-EJ known as polymerase theta dependent end joining. Finally, whole genome sequencing analysis showed HPV8 E6 caused an increased frequency of deletions bearing the microhomology signatures of Alt-EJ. This study fills the knowledge gap of how DSB is repaired in cells with HPV8 E6 and the mutagenic consequences of HPV8 E6 mediated p300 destabilization. Broadly, this study supports the hypothesis that beta-HPV promotes cancer formation by increasing genomic instability.

Data availability

Sequences have been deposited in the NCBI SRA database with accession number (PRJNA 856469).

The following data sets were generated

Article and author information

Author details

  1. Changkun Hu

    Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4407-7144
  2. Taylor Bugbee

    Division of Biology, Kansas State University, Manhattan, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Rachel Palinski

    Veterinary Diagnostic Laboratory, Kansas State University, Manhattan, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Ibukun A Akinyemi

    Department of Pediatrics, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Michael T McIntosh

    Department of Pediatrics, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Thomas MacCarthy

    Laufer Center for Physical and Quantitative Biology, Stony Brook University, Stony Brook, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Sumita Bhaduri-McIntosh

    Department of Pediatrics, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Nicholas Wallace

    Division of Biology, Kansas State University, Manhattan, United States
    For correspondence
    nwallac@ksu.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3971-716X

Funding

National Institute of General Medical Sciences (P20GM130448)

  • Nicholas Wallace

NIH Research Enhancement Award (NCI R15 CA242057 01A1)

  • Nicholas Wallace

U.S. Department of Defense (CMDRP PRCRP CA160224 (NW))

  • Nicholas Wallace

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2023, Hu et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,161
    views
  • 140
    downloads
  • 5
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Changkun Hu
  2. Taylor Bugbee
  3. Rachel Palinski
  4. Ibukun A Akinyemi
  5. Michael T McIntosh
  6. Thomas MacCarthy
  7. Sumita Bhaduri-McIntosh
  8. Nicholas Wallace
(2023)
Beta human papillomavirus 8E6 promotes alternative end-joining
eLife 12:e81923.
https://doi.org/10.7554/eLife.81923

Share this article

https://doi.org/10.7554/eLife.81923

Further reading

    1. Cancer Biology
    2. Physics of Living Systems
    Joseph Ackermann, Chiara Bernard ... Martine D Ben Amar
    Research Article

    The tumor stroma consists mainly of extracellular matrix, fibroblasts, immune cells, and vasculature. Its structure and functions are altered during malignancy: tumor cells transform fibroblasts into cancer-associated fibroblasts, which exhibit immunosuppressive activities on which growth and metastasis depend. These include exclusion of immune cells from the tumor nest, cancer progression, and inhibition of T-cell-based immunotherapy. To understand these complex interactions, we measure the density of different cell types in the stroma using immunohistochemistry techniques on tumor samples from lung cancer patients. We incorporate these data into a minimal dynamical system, explore the variety of outcomes, and finally establish a spatio-temporal model that explains the cell distribution. We reproduce that cancer-associated fibroblasts act as a barrier to tumor expansion, but also reduce the efficiency of the immune response. Our conclusion is that the final outcome depends on the parameter values for each patient and leads to either tumor invasion, persistence, or eradication as a result of the interplay between cancer cell growth, T-cell cytotoxicity, and fibroblast activity. However, despite the existence of a wide range of scenarios, distinct trajectories, and patterns allow quantitative predictions that may help in the selection of new therapies and personalized protocols.

    1. Cancer Biology
    Huan Fang, Huichun Liang ... Ceshi Chen
    Research Article

    In the clinic, anti-tumor angiogenesis is commonly employed for treating recurrent, metastatic, drug-resistant triple-negative, and advanced breast cancer. Our previous research revealed that the deubiquitinase STAMBPL1 enhances the stability of MKP-1, thereby promoting cisplatin resistance in breast cancer. In this study, we discovered that STAMBPL1 could upregulate the expression of the hypoxia-inducible factor HIF1α in breast cancer cells. Therefore, we investigated whether STAMBPL1 promotes tumor angiogenesis. We demonstrated that STAMBPL1 increased HIF1A transcription in a non-enzymatic manner, thereby activating the HIF1α/VEGFA signaling pathway to facilitate triple-negative breast cancer angiogenesis. Through RNA-seq analysis, we identified the transcription factor GRHL3 as a downstream target of STAMBPL1 that is responsible for mediating HIF1A transcription. Furthermore, we discovered that STAMBPL1 regulates GRHL3 transcription by interacting with the transcription factor FOXO1. These findings shed light on the role and mechanism of STAMBPL1 in the pathogenesis of breast cancer, offering novel targets and avenues for the treatment of triple-negative and advanced breast cancer.