Argonaute 4 as an Effector Protein in RNA-Directed DNA Methylation in Human Cells
DNA methylation of specific genome locations contributes to the distinct functions of multicellular organisms. DNA methylation can be governed by RNA-dependent DNA methylation (RdDM). RdDM is carried out by endogenous small-RNA-guided epigenomic editing complexes that add a methyl group to a precise...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-07-01
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| Series: | Frontiers in Genetics |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fgene.2019.00645/full |
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| author | Kanwalat Chalertpet Piyapat Pin-on Piyapat Pin-on Chatchawit Aporntewan Maturada Patchsung Maturada Patchsung Praewphan Ingrungruanglert Nipan Israsena Apiwat Mutirangura Apiwat Mutirangura |
| author_facet | Kanwalat Chalertpet Piyapat Pin-on Piyapat Pin-on Chatchawit Aporntewan Maturada Patchsung Maturada Patchsung Praewphan Ingrungruanglert Nipan Israsena Apiwat Mutirangura Apiwat Mutirangura |
| author_sort | Kanwalat Chalertpet |
| collection | DOAJ |
| description | DNA methylation of specific genome locations contributes to the distinct functions of multicellular organisms. DNA methylation can be governed by RNA-dependent DNA methylation (RdDM). RdDM is carried out by endogenous small-RNA-guided epigenomic editing complexes that add a methyl group to a precise DNA location. In plants, the Argonaute 4 (AGO4) protein is one of the main catalytic components involved in RdDM. Although small interfering RNA or short hairpin RNA has been shown to be able to guide DNA methylation in human cells, AGO protein-regulated RdDM in humans has not yet been evaluated. This study aimed to identify a key regulatory AGO protein involved in human RdDM by bioinformatics and to explore its function in RdDM by a combination of AGO4 knockdown, Alu small interfering RNA transfection, AGO4-expressing plasmid transfection, chromatin immunoprecipitation, cell-penetrating peptide-tagged AGO4 combined Alu single-guide RNA transfection, and methylation analyses. We found that first, human AGO4 showed stronger genome-wide association with DNA methylation than AGO1–AGO3. Second, endogenous AGO4 depletion demethylated DNA of known AGO4 bound loci. Finally, exogenous AGO4 de novo methylated the bound DNA sequences. Therefore, we discovered that AGO4 plays a role in human RdDM. |
| first_indexed | 2024-12-11T16:46:44Z |
| format | Article |
| id | doaj.art-e34aeeb783144780b1f849ed087caa6f |
| institution | Directory Open Access Journal |
| issn | 1664-8021 |
| language | English |
| last_indexed | 2024-12-11T16:46:44Z |
| publishDate | 2019-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Genetics |
| spelling | doaj.art-e34aeeb783144780b1f849ed087caa6f2022-12-22T00:58:11ZengFrontiers Media S.A.Frontiers in Genetics1664-80212019-07-011010.3389/fgene.2019.00645463961Argonaute 4 as an Effector Protein in RNA-Directed DNA Methylation in Human CellsKanwalat Chalertpet0Piyapat Pin-on1Piyapat Pin-on2Chatchawit Aporntewan3Maturada Patchsung4Maturada Patchsung5Praewphan Ingrungruanglert6Nipan Israsena7Apiwat Mutirangura8Apiwat Mutirangura9Interdisciplinary Program of Biomedical Sciences, Faculty of the Graduate School, Chulalongkorn University, Bangkok, ThailandInterdisciplinary Program of Biomedical Sciences, Faculty of the Graduate School, Chulalongkorn University, Bangkok, ThailandFaculty of Medicine, King Mongkut’s Institute of Technology Ladkrabang, Bangkok, ThailandDepartment of Mathematics and Computer Science, Faculty of Science, Chulalongkorn University, Bangkok, ThailandInterdisciplinary Program of Biomedical Sciences, Faculty of the Graduate School, Chulalongkorn University, Bangkok, ThailandCenter of Excellence in Molecular Genetics of Cancer and Human Diseases, Department of Anatomy, Faculty of Medicine, Chulalongkorn University, Bangkok, ThailandStem Cells and Cell Therapy Research Unit, Faculty of Medicine, Chulalongkorn University, Bangkok, ThailandStem Cells and Cell Therapy Research Unit, Faculty of Medicine, Chulalongkorn University, Bangkok, ThailandCenter of Excellence in Molecular Genetics of Cancer and Human Diseases, Department of Anatomy, Faculty of Medicine, Chulalongkorn University, Bangkok, ThailandDepartment of Anatomy, Faculty of Medicine, Chulalongkorn University, Bangkok, ThailandDNA methylation of specific genome locations contributes to the distinct functions of multicellular organisms. DNA methylation can be governed by RNA-dependent DNA methylation (RdDM). RdDM is carried out by endogenous small-RNA-guided epigenomic editing complexes that add a methyl group to a precise DNA location. In plants, the Argonaute 4 (AGO4) protein is one of the main catalytic components involved in RdDM. Although small interfering RNA or short hairpin RNA has been shown to be able to guide DNA methylation in human cells, AGO protein-regulated RdDM in humans has not yet been evaluated. This study aimed to identify a key regulatory AGO protein involved in human RdDM by bioinformatics and to explore its function in RdDM by a combination of AGO4 knockdown, Alu small interfering RNA transfection, AGO4-expressing plasmid transfection, chromatin immunoprecipitation, cell-penetrating peptide-tagged AGO4 combined Alu single-guide RNA transfection, and methylation analyses. We found that first, human AGO4 showed stronger genome-wide association with DNA methylation than AGO1–AGO3. Second, endogenous AGO4 depletion demethylated DNA of known AGO4 bound loci. Finally, exogenous AGO4 de novo methylated the bound DNA sequences. Therefore, we discovered that AGO4 plays a role in human RdDM.https://www.frontiersin.org/article/10.3389/fgene.2019.00645/fullhuman Argonaute 4human RNA-directed DNA methylationhuman RdDMCPP-AGO4epigenomic editing |
| spellingShingle | Kanwalat Chalertpet Piyapat Pin-on Piyapat Pin-on Chatchawit Aporntewan Maturada Patchsung Maturada Patchsung Praewphan Ingrungruanglert Nipan Israsena Apiwat Mutirangura Apiwat Mutirangura Argonaute 4 as an Effector Protein in RNA-Directed DNA Methylation in Human Cells Frontiers in Genetics human Argonaute 4 human RNA-directed DNA methylation human RdDM CPP-AGO4 epigenomic editing |
| title | Argonaute 4 as an Effector Protein in RNA-Directed DNA Methylation in Human Cells |
| title_full | Argonaute 4 as an Effector Protein in RNA-Directed DNA Methylation in Human Cells |
| title_fullStr | Argonaute 4 as an Effector Protein in RNA-Directed DNA Methylation in Human Cells |
| title_full_unstemmed | Argonaute 4 as an Effector Protein in RNA-Directed DNA Methylation in Human Cells |
| title_short | Argonaute 4 as an Effector Protein in RNA-Directed DNA Methylation in Human Cells |
| title_sort | argonaute 4 as an effector protein in rna directed dna methylation in human cells |
| topic | human Argonaute 4 human RNA-directed DNA methylation human RdDM CPP-AGO4 epigenomic editing |
| url | https://www.frontiersin.org/article/10.3389/fgene.2019.00645/full |
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