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Genome-Wide CRISPR Screens Reveal ZATT as a Synthetic Lethal Target of TOP2-Poison Etoposide That Can Act in a TDP2-Independent Pathway

Etoposide (ETO) is an anticancer drug that targets topoisomerase II (TOP2). It stabilizes a normally transient TOP2–DNA covalent complex (TOP2cc), thus leading to DNA double-strand breaks (DSBs). Tyrosyl-DNA phosphodiesterases two (TDP2) is directly involved in the repair of TOP2cc by removing phosp...

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Autores principales: Park, Jeong-Min, Zhang, Huimin, Nie, Litong, Wang, Chao, Huang, Min, Feng, Xu, Tang, Mengfan, Chen, Zhen, Xiong, Yun, Lee, Namsoo, Li, Siting, Yin, Ling, Hart, Traver, Chen, Junjie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10095316/
https://www.ncbi.nlm.nih.gov/pubmed/37047518
http://dx.doi.org/10.3390/ijms24076545
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author Park, Jeong-Min
Zhang, Huimin
Nie, Litong
Wang, Chao
Huang, Min
Feng, Xu
Tang, Mengfan
Chen, Zhen
Xiong, Yun
Lee, Namsoo
Li, Siting
Yin, Ling
Hart, Traver
Chen, Junjie
author_facet Park, Jeong-Min
Zhang, Huimin
Nie, Litong
Wang, Chao
Huang, Min
Feng, Xu
Tang, Mengfan
Chen, Zhen
Xiong, Yun
Lee, Namsoo
Li, Siting
Yin, Ling
Hart, Traver
Chen, Junjie
author_sort Park, Jeong-Min
collection PubMed
description Etoposide (ETO) is an anticancer drug that targets topoisomerase II (TOP2). It stabilizes a normally transient TOP2–DNA covalent complex (TOP2cc), thus leading to DNA double-strand breaks (DSBs). Tyrosyl-DNA phosphodiesterases two (TDP2) is directly involved in the repair of TOP2cc by removing phosphotyrosyl peptides from 5′-termini of DSBs. Recent studies suggest that additional factors are required for TOP2cc repair, which include the proteasome and the zinc finger protein associated with TDP2 and TOP2, named ZATT. ZATT may alter the conformation of TOP2cc in a way that renders the accessibility of TDP2 for TOP2cc removal. In this study, our genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) screens revealed that ZATT also has a TDP2-independent role in promoting cell survival following ETO treatment. ZATT KO cells showed relatively higher ETO sensitivity than TDP2-KO cells, and ZATT/TDP2 DKO cells displayed additive hypersensitivity to ETO treatment. The study using a series of deletion mutants of ZATT determined that the N-terminal 1–168 residues of ZATT are required for interaction with TOP2 and this interaction is critical to ETO sensitivity. Moreover, depletion of ZATT resulted in accelerated TOP2 degradation after ETO or cycloheximide (CHX) treatment, suggesting that ZATT may increase TOP2 stability and likely participate in TOP2 turnover. Taken together, this study suggests that ZATT is a critical determinant that dictates responses to ETO treatment and targeting. ZATT is a promising strategy to increase ETO efficacy for cancer therapy.
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spelling pubmed-100953162023-04-13 Genome-Wide CRISPR Screens Reveal ZATT as a Synthetic Lethal Target of TOP2-Poison Etoposide That Can Act in a TDP2-Independent Pathway Park, Jeong-Min Zhang, Huimin Nie, Litong Wang, Chao Huang, Min Feng, Xu Tang, Mengfan Chen, Zhen Xiong, Yun Lee, Namsoo Li, Siting Yin, Ling Hart, Traver Chen, Junjie Int J Mol Sci Article Etoposide (ETO) is an anticancer drug that targets topoisomerase II (TOP2). It stabilizes a normally transient TOP2–DNA covalent complex (TOP2cc), thus leading to DNA double-strand breaks (DSBs). Tyrosyl-DNA phosphodiesterases two (TDP2) is directly involved in the repair of TOP2cc by removing phosphotyrosyl peptides from 5′-termini of DSBs. Recent studies suggest that additional factors are required for TOP2cc repair, which include the proteasome and the zinc finger protein associated with TDP2 and TOP2, named ZATT. ZATT may alter the conformation of TOP2cc in a way that renders the accessibility of TDP2 for TOP2cc removal. In this study, our genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) screens revealed that ZATT also has a TDP2-independent role in promoting cell survival following ETO treatment. ZATT KO cells showed relatively higher ETO sensitivity than TDP2-KO cells, and ZATT/TDP2 DKO cells displayed additive hypersensitivity to ETO treatment. The study using a series of deletion mutants of ZATT determined that the N-terminal 1–168 residues of ZATT are required for interaction with TOP2 and this interaction is critical to ETO sensitivity. Moreover, depletion of ZATT resulted in accelerated TOP2 degradation after ETO or cycloheximide (CHX) treatment, suggesting that ZATT may increase TOP2 stability and likely participate in TOP2 turnover. Taken together, this study suggests that ZATT is a critical determinant that dictates responses to ETO treatment and targeting. ZATT is a promising strategy to increase ETO efficacy for cancer therapy. MDPI 2023-03-31 /pmc/articles/PMC10095316/ /pubmed/37047518 http://dx.doi.org/10.3390/ijms24076545 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Park, Jeong-Min
Zhang, Huimin
Nie, Litong
Wang, Chao
Huang, Min
Feng, Xu
Tang, Mengfan
Chen, Zhen
Xiong, Yun
Lee, Namsoo
Li, Siting
Yin, Ling
Hart, Traver
Chen, Junjie
Genome-Wide CRISPR Screens Reveal ZATT as a Synthetic Lethal Target of TOP2-Poison Etoposide That Can Act in a TDP2-Independent Pathway
title Genome-Wide CRISPR Screens Reveal ZATT as a Synthetic Lethal Target of TOP2-Poison Etoposide That Can Act in a TDP2-Independent Pathway
title_full Genome-Wide CRISPR Screens Reveal ZATT as a Synthetic Lethal Target of TOP2-Poison Etoposide That Can Act in a TDP2-Independent Pathway
title_fullStr Genome-Wide CRISPR Screens Reveal ZATT as a Synthetic Lethal Target of TOP2-Poison Etoposide That Can Act in a TDP2-Independent Pathway
title_full_unstemmed Genome-Wide CRISPR Screens Reveal ZATT as a Synthetic Lethal Target of TOP2-Poison Etoposide That Can Act in a TDP2-Independent Pathway
title_short Genome-Wide CRISPR Screens Reveal ZATT as a Synthetic Lethal Target of TOP2-Poison Etoposide That Can Act in a TDP2-Independent Pathway
title_sort genome-wide crispr screens reveal zatt as a synthetic lethal target of top2-poison etoposide that can act in a tdp2-independent pathway
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10095316/
https://www.ncbi.nlm.nih.gov/pubmed/37047518
http://dx.doi.org/10.3390/ijms24076545
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