DNA Damage Induction Alters the Expression of Ubiquitin and SUMO Regulators in Preimplantation Stage Pig Embryos
DNA damage in early-stage embryos impacts development and is a risk factor for segregation of altered genomes. DNA damage response (DDR) encompasses a sophisticated network of proteins involved in sensing, signaling, and repairing damage. DDR is regulated by reversible post-translational modificatio...
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| Format: | Article |
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MDPI AG
2022-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/17/9610 |
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| author | Zigomar da Silva Werner Giehl Glanzner Luke Currin Mariana Priotto de Macedo Karina Gutierrez Vanessa Guay Paulo Bayard Dias Gonçalves Vilceu Bordignon |
| author_facet | Zigomar da Silva Werner Giehl Glanzner Luke Currin Mariana Priotto de Macedo Karina Gutierrez Vanessa Guay Paulo Bayard Dias Gonçalves Vilceu Bordignon |
| author_sort | Zigomar da Silva |
| collection | DOAJ |
| description | DNA damage in early-stage embryos impacts development and is a risk factor for segregation of altered genomes. DNA damage response (DDR) encompasses a sophisticated network of proteins involved in sensing, signaling, and repairing damage. DDR is regulated by reversible post-translational modifications including acetylation, methylation, phosphorylation, ubiquitylation, and SUMOylation. While important regulators of these processes have been characterized in somatic cells, their roles in early-stage embryos remain broadly unknown. The objective of this study was to explore how ubiquitylation and SUMOylation are involved in the regulation of early development in porcine embryos by assessing the mRNA profile of genes encoding ubiquitination (UBs), deubiquitination (DUBs), SUMOylation (SUMOs) or deSUMOylation (deSUMOs) enzymes in oocyte and embryos at different stages of development, and to evaluate if the induction of DNA damage at different stages of embryo development would alter the mRNA abundance of these genes. Pig embryos were produced by in vitro fertilization and DNA damage was induced by ultraviolet (UV) light exposure for 10 s on days 2, 4 or 7 of development. The relative mRNA abundance of most UBs, DUBs, SUMOs, and deSUMOs was higher in oocytes and early-stage embryos than in blastocysts. Transcript levels for UBs (<i>RNF20</i>, <i>RNF40</i>, <i>RNF114</i>, <i>RNF169</i>, <i>CUL5</i>, <i>DCAF2</i>, <i>DECAF13</i>, and <i>DDB1</i>), DUBs (<i>USP16</i>), and SUMOs (<i>CBX4</i>, <i>UBA2</i> and <i>UBC9</i>), were upregulated in early-stage embryos (D2 and/or D4) compared to oocytes and blastocysts. In response to UV-induced DNA damage, transcript levels of several UBs, DUBs, SUMOs, and deSUMOs decreased in D2 and D4 embryos, but increased in blastocysts. These findings revealed that transcript levels of genes encoding for important UBs, DUBs, SUMOs, and deSUMOs are regulated during early embryo development and are modulated in response to induced DNA damage. This study has also identified candidate genes controlling post-translational modifications that may have relevant roles in the regulation of normal embryo development, repair of damaged DNA, and preservation of genome stability in the pig embryo. |
| first_indexed | 2024-03-10T01:46:05Z |
| format | Article |
| id | doaj.art-45cd0a8c498042338a87f11998347bb4 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T01:46:05Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-45cd0a8c498042338a87f11998347bb42023-11-23T13:13:57ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-08-012317961010.3390/ijms23179610DNA Damage Induction Alters the Expression of Ubiquitin and SUMO Regulators in Preimplantation Stage Pig EmbryosZigomar da Silva0Werner Giehl Glanzner1Luke Currin2Mariana Priotto de Macedo3Karina Gutierrez4Vanessa Guay5Paulo Bayard Dias Gonçalves6Vilceu Bordignon7Laboratory of Biotechnology and Animal Reproduction–BioRep, Federal University of Santa Maria, Santa Maria 97105-900, BrazilDepartment of Animal Science, McGill University, Sainte-Anne-de-Bellevue, QC H9X 3V9, CanadaDepartment of Animal Science, McGill University, Sainte-Anne-de-Bellevue, QC H9X 3V9, CanadaDepartment of Animal Science, McGill University, Sainte-Anne-de-Bellevue, QC H9X 3V9, CanadaDepartment of Animal Science, McGill University, Sainte-Anne-de-Bellevue, QC H9X 3V9, CanadaDepartment of Animal Science, McGill University, Sainte-Anne-de-Bellevue, QC H9X 3V9, CanadaLaboratory of Biotechnology and Animal Reproduction–BioRep, Federal University of Santa Maria, Santa Maria 97105-900, BrazilDepartment of Animal Science, McGill University, Sainte-Anne-de-Bellevue, QC H9X 3V9, CanadaDNA damage in early-stage embryos impacts development and is a risk factor for segregation of altered genomes. DNA damage response (DDR) encompasses a sophisticated network of proteins involved in sensing, signaling, and repairing damage. DDR is regulated by reversible post-translational modifications including acetylation, methylation, phosphorylation, ubiquitylation, and SUMOylation. While important regulators of these processes have been characterized in somatic cells, their roles in early-stage embryos remain broadly unknown. The objective of this study was to explore how ubiquitylation and SUMOylation are involved in the regulation of early development in porcine embryos by assessing the mRNA profile of genes encoding ubiquitination (UBs), deubiquitination (DUBs), SUMOylation (SUMOs) or deSUMOylation (deSUMOs) enzymes in oocyte and embryos at different stages of development, and to evaluate if the induction of DNA damage at different stages of embryo development would alter the mRNA abundance of these genes. Pig embryos were produced by in vitro fertilization and DNA damage was induced by ultraviolet (UV) light exposure for 10 s on days 2, 4 or 7 of development. The relative mRNA abundance of most UBs, DUBs, SUMOs, and deSUMOs was higher in oocytes and early-stage embryos than in blastocysts. Transcript levels for UBs (<i>RNF20</i>, <i>RNF40</i>, <i>RNF114</i>, <i>RNF169</i>, <i>CUL5</i>, <i>DCAF2</i>, <i>DECAF13</i>, and <i>DDB1</i>), DUBs (<i>USP16</i>), and SUMOs (<i>CBX4</i>, <i>UBA2</i> and <i>UBC9</i>), were upregulated in early-stage embryos (D2 and/or D4) compared to oocytes and blastocysts. In response to UV-induced DNA damage, transcript levels of several UBs, DUBs, SUMOs, and deSUMOs decreased in D2 and D4 embryos, but increased in blastocysts. These findings revealed that transcript levels of genes encoding for important UBs, DUBs, SUMOs, and deSUMOs are regulated during early embryo development and are modulated in response to induced DNA damage. This study has also identified candidate genes controlling post-translational modifications that may have relevant roles in the regulation of normal embryo development, repair of damaged DNA, and preservation of genome stability in the pig embryo.https://www.mdpi.com/1422-0067/23/17/9610swineembryo developmentubiquitylationSUMOylationDNA damage |
| spellingShingle | Zigomar da Silva Werner Giehl Glanzner Luke Currin Mariana Priotto de Macedo Karina Gutierrez Vanessa Guay Paulo Bayard Dias Gonçalves Vilceu Bordignon DNA Damage Induction Alters the Expression of Ubiquitin and SUMO Regulators in Preimplantation Stage Pig Embryos International Journal of Molecular Sciences swine embryo development ubiquitylation SUMOylation DNA damage |
| title | DNA Damage Induction Alters the Expression of Ubiquitin and SUMO Regulators in Preimplantation Stage Pig Embryos |
| title_full | DNA Damage Induction Alters the Expression of Ubiquitin and SUMO Regulators in Preimplantation Stage Pig Embryos |
| title_fullStr | DNA Damage Induction Alters the Expression of Ubiquitin and SUMO Regulators in Preimplantation Stage Pig Embryos |
| title_full_unstemmed | DNA Damage Induction Alters the Expression of Ubiquitin and SUMO Regulators in Preimplantation Stage Pig Embryos |
| title_short | DNA Damage Induction Alters the Expression of Ubiquitin and SUMO Regulators in Preimplantation Stage Pig Embryos |
| title_sort | dna damage induction alters the expression of ubiquitin and sumo regulators in preimplantation stage pig embryos |
| topic | swine embryo development ubiquitylation SUMOylation DNA damage |
| url | https://www.mdpi.com/1422-0067/23/17/9610 |
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