Beta human papillomavirus 8E6 promotes alternative end joining
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 repa...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2023-01-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/81923 |
| _version_ | 1811172344953044992 |
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| author | Changkun Hu Taylor Bugbee Rachel Palinski Ibukun A Akinyemi Michael T McIntosh Thomas MacCarthy Sumita Bhaduri-McIntosh Nicholas Wallace |
| author_facet | Changkun Hu Taylor Bugbee Rachel Palinski Ibukun A Akinyemi Michael T McIntosh Thomas MacCarthy Sumita Bhaduri-McIntosh Nicholas Wallace |
| author_sort | Changkun Hu |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-10T17:28:26Z |
| format | Article |
| id | doaj.art-b54c18b5addc4591aac934765ee5ce95 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-10T17:28:26Z |
| publishDate | 2023-01-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-b54c18b5addc4591aac934765ee5ce952023-02-03T16:58:48ZengeLife Sciences Publications LtdeLife2050-084X2023-01-011210.7554/eLife.81923Beta human papillomavirus 8E6 promotes alternative end joiningChangkun Hu0https://orcid.org/0000-0002-4407-7144Taylor Bugbee1Rachel Palinski2Ibukun A Akinyemi3Michael T McIntosh4Thomas MacCarthy5Sumita Bhaduri-McIntosh6Nicholas Wallace7https://orcid.org/0000-0002-3971-716XBasic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, United States; Division of Biology, Kansas State University, Manhattan, United StatesDivision of Biology, Kansas State University, Manhattan, United StatesVeterinary Diagnostic Laboratory, Kansas State University, Manhattan, United StatesChild Health Research Institute, Department of Pediatrics, University of Florida, Gainesville, United States; Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, United StatesChild Health Research Institute, Department of Pediatrics, University of Florida, Gainesville, United StatesLaufer Center for Physical and Quantitative Biology, Stony Brook University, Stony Brook, United StatesChild Health Research Institute, Department of Pediatrics, University of Florida, Gainesville, United States; Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, United StatesDivision of Biology, Kansas State University, Manhattan, United StatesDouble 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.https://elifesciences.org/articles/81923beta-HPVDSB repairMMEJP300NHEJHR |
| spellingShingle | Changkun Hu Taylor Bugbee Rachel Palinski Ibukun A Akinyemi Michael T McIntosh Thomas MacCarthy Sumita Bhaduri-McIntosh Nicholas Wallace Beta human papillomavirus 8E6 promotes alternative end joining eLife beta-HPV DSB repair MMEJ P300 NHEJ HR |
| title | Beta human papillomavirus 8E6 promotes alternative end joining |
| title_full | Beta human papillomavirus 8E6 promotes alternative end joining |
| title_fullStr | Beta human papillomavirus 8E6 promotes alternative end joining |
| title_full_unstemmed | Beta human papillomavirus 8E6 promotes alternative end joining |
| title_short | Beta human papillomavirus 8E6 promotes alternative end joining |
| title_sort | beta human papillomavirus 8e6 promotes alternative end joining |
| topic | beta-HPV DSB repair MMEJ P300 NHEJ HR |
| url | https://elifesciences.org/articles/81923 |
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