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Similar deamination activities but different phenotypic outcomes induced by APOBEC3 enzymes in breast epithelial cells
APOBEC3 (A3) enzymes deaminate cytosine to uracil in viral single-stranded DNA as a mutagenic barrier for some viruses. A3-induced deaminations can also occur in human genomes resulting in an endogenous source of somatic mutations in multiple cancers. However, the roles of each A3 are unclear since...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10267419/ https://www.ncbi.nlm.nih.gov/pubmed/37324648 http://dx.doi.org/10.3389/fgeed.2023.1196697 |
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author | Granadillo Rodríguez, Milaid Wong, Lai Chelico, Linda |
author_facet | Granadillo Rodríguez, Milaid Wong, Lai Chelico, Linda |
author_sort | Granadillo Rodríguez, Milaid |
collection | PubMed |
description | APOBEC3 (A3) enzymes deaminate cytosine to uracil in viral single-stranded DNA as a mutagenic barrier for some viruses. A3-induced deaminations can also occur in human genomes resulting in an endogenous source of somatic mutations in multiple cancers. However, the roles of each A3 are unclear since few studies have assessed these enzymes in parallel. Thus, we developed stable cell lines expressing A3A, A3B, or A3H Hap I using non-tumorigenic MCF10A and tumorigenic MCF7 breast epithelial cells to assess their mutagenic potential and cancer phenotypes in breast cells. The activity of these enzymes was characterized by γH2AX foci formation and in vitro deamination. Cell migration and soft agar colony formation assays assessed cellular transformation potential. We found that all three A3 enzymes had similar γH2AX foci formation, despite different deamination activities in vitro. Notably, in nuclear lysates, the in vitro deaminase activity of A3A, A3B, and A3H did not require digestion of cellular RNA, in contrast to that of A3B and A3H in whole-cell lysates. Their similar activities in cells, nonetheless, resulted in distinct phenotypes where A3A decreased colony formation in soft agar, A3B decreased colony formation in soft agar after hydroxyurea treatment, and A3H Hap I promoted cell migration. Overall, we show that in vitro deamination data do not always reflect cell DNA damage, all three A3s induce DNA damage, and the impact of each is different. |
format | Online Article Text |
id | pubmed-10267419 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-102674192023-06-15 Similar deamination activities but different phenotypic outcomes induced by APOBEC3 enzymes in breast epithelial cells Granadillo Rodríguez, Milaid Wong, Lai Chelico, Linda Front Genome Ed Genome Editing APOBEC3 (A3) enzymes deaminate cytosine to uracil in viral single-stranded DNA as a mutagenic barrier for some viruses. A3-induced deaminations can also occur in human genomes resulting in an endogenous source of somatic mutations in multiple cancers. However, the roles of each A3 are unclear since few studies have assessed these enzymes in parallel. Thus, we developed stable cell lines expressing A3A, A3B, or A3H Hap I using non-tumorigenic MCF10A and tumorigenic MCF7 breast epithelial cells to assess their mutagenic potential and cancer phenotypes in breast cells. The activity of these enzymes was characterized by γH2AX foci formation and in vitro deamination. Cell migration and soft agar colony formation assays assessed cellular transformation potential. We found that all three A3 enzymes had similar γH2AX foci formation, despite different deamination activities in vitro. Notably, in nuclear lysates, the in vitro deaminase activity of A3A, A3B, and A3H did not require digestion of cellular RNA, in contrast to that of A3B and A3H in whole-cell lysates. Their similar activities in cells, nonetheless, resulted in distinct phenotypes where A3A decreased colony formation in soft agar, A3B decreased colony formation in soft agar after hydroxyurea treatment, and A3H Hap I promoted cell migration. Overall, we show that in vitro deamination data do not always reflect cell DNA damage, all three A3s induce DNA damage, and the impact of each is different. Frontiers Media S.A. 2023-06-01 /pmc/articles/PMC10267419/ /pubmed/37324648 http://dx.doi.org/10.3389/fgeed.2023.1196697 Text en Copyright © 2023 Granadillo Rodríguez, Wong and Chelico. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Genome Editing Granadillo Rodríguez, Milaid Wong, Lai Chelico, Linda Similar deamination activities but different phenotypic outcomes induced by APOBEC3 enzymes in breast epithelial cells |
title | Similar deamination activities but different phenotypic outcomes induced by APOBEC3 enzymes in breast epithelial cells |
title_full | Similar deamination activities but different phenotypic outcomes induced by APOBEC3 enzymes in breast epithelial cells |
title_fullStr | Similar deamination activities but different phenotypic outcomes induced by APOBEC3 enzymes in breast epithelial cells |
title_full_unstemmed | Similar deamination activities but different phenotypic outcomes induced by APOBEC3 enzymes in breast epithelial cells |
title_short | Similar deamination activities but different phenotypic outcomes induced by APOBEC3 enzymes in breast epithelial cells |
title_sort | similar deamination activities but different phenotypic outcomes induced by apobec3 enzymes in breast epithelial cells |
topic | Genome Editing |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10267419/ https://www.ncbi.nlm.nih.gov/pubmed/37324648 http://dx.doi.org/10.3389/fgeed.2023.1196697 |
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