UAP56 associates with DRM2 and is localized to chromatin in Arabidopsis
Repeated sequence expression and transposable element mobilization are tightly controlled by multilayer processes, which include DNA 5′‐cytosine methylation. The RNA‐directed DNA methylation (RdDM) pathway, which uses siRNAs to guide sequence‐specific directed DNA methylation, emerged specifically i...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2019-05-01
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| Series: | FEBS Open Bio |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/2211-5463.12627 |
| _version_ | 1798018780518940672 |
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| author | Jacinthe Azevedo Claire Picart Laurent Dureau Dominique Pontier Sylvie Jaquinod‐Kieffer Mohamed‐Ali Hakimi Thierry Lagrange |
| author_facet | Jacinthe Azevedo Claire Picart Laurent Dureau Dominique Pontier Sylvie Jaquinod‐Kieffer Mohamed‐Ali Hakimi Thierry Lagrange |
| author_sort | Jacinthe Azevedo |
| collection | DOAJ |
| description | Repeated sequence expression and transposable element mobilization are tightly controlled by multilayer processes, which include DNA 5′‐cytosine methylation. The RNA‐directed DNA methylation (RdDM) pathway, which uses siRNAs to guide sequence‐specific directed DNA methylation, emerged specifically in plants. RdDM ensures DNA methylation maintenance on asymmetric CHH sites and specifically initiates de novo methylation in all cytosine sequence contexts through the action of DRM DNA methyltransferases, of which DRM2 is the most prominent. The RdDM pathway has been well described, but how DRM2 is recruited onto DNA targets and associates with other RdDM factors remains unknown. To address these questions, we developed biochemical approaches to allow the identification of factors that may escape genetic screens, such as proteins encoded by multigenic families. Through both conventional and affinity purification of DRM2, we identified DEAD box RNA helicases U2AF56 Associated Protein 56 (UAP56a/b), which are widespread among eukaryotes, as new DRM2 partners. We have shown that, similar to DRM2 and other RdDM actors, UAP56 has chromatin‐associated protein properties. We confirmed this association both in vitro and in vivo in reproductive tissues. In addition, our experiments also suggest that UAP56 may exhibit differential distribution in cells depending on plant organ. While originally identified for its role in splicing, our study suggests that UAP56 may also have other roles, and our findings allow us to initiate discussion about its potential role in the RdDM pathway. |
| first_indexed | 2024-04-11T16:29:53Z |
| format | Article |
| id | doaj.art-033f1a6f81bc4b5384af7dbf5918f62c |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-04-11T16:29:53Z |
| publishDate | 2019-05-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-033f1a6f81bc4b5384af7dbf5918f62c2022-12-22T04:14:04ZengWileyFEBS Open Bio2211-54632019-05-019597398510.1002/2211-5463.12627UAP56 associates with DRM2 and is localized to chromatin in ArabidopsisJacinthe Azevedo0Claire Picart1Laurent Dureau2Dominique Pontier3Sylvie Jaquinod‐Kieffer4Mohamed‐Ali Hakimi5Thierry Lagrange6LGDP‐UMR5096 CNRS Perpignan FranceLGDP‐UMR5096 CNRS Perpignan FranceLGDP‐UMR5096 CNRS Perpignan FranceLGDP‐UMR5096 CNRS Perpignan FranceLaboratoire Biologie Grande Echelle Institut de Biosciences et Biotechnologies de Grenoble UMR_S 1038 CEA, INSERM Université Grenoble Alpes FranceInstitute for Advanced Biosciences (IAB) Team Host‐pathogen Interactions and Immunity to Infection INSERM U1209 CNRS UMR5309 Université Grenoble Alpes FranceLGDP‐UMR5096 CNRS Perpignan FranceRepeated sequence expression and transposable element mobilization are tightly controlled by multilayer processes, which include DNA 5′‐cytosine methylation. The RNA‐directed DNA methylation (RdDM) pathway, which uses siRNAs to guide sequence‐specific directed DNA methylation, emerged specifically in plants. RdDM ensures DNA methylation maintenance on asymmetric CHH sites and specifically initiates de novo methylation in all cytosine sequence contexts through the action of DRM DNA methyltransferases, of which DRM2 is the most prominent. The RdDM pathway has been well described, but how DRM2 is recruited onto DNA targets and associates with other RdDM factors remains unknown. To address these questions, we developed biochemical approaches to allow the identification of factors that may escape genetic screens, such as proteins encoded by multigenic families. Through both conventional and affinity purification of DRM2, we identified DEAD box RNA helicases U2AF56 Associated Protein 56 (UAP56a/b), which are widespread among eukaryotes, as new DRM2 partners. We have shown that, similar to DRM2 and other RdDM actors, UAP56 has chromatin‐associated protein properties. We confirmed this association both in vitro and in vivo in reproductive tissues. In addition, our experiments also suggest that UAP56 may exhibit differential distribution in cells depending on plant organ. While originally identified for its role in splicing, our study suggests that UAP56 may also have other roles, and our findings allow us to initiate discussion about its potential role in the RdDM pathway.https://doi.org/10.1002/2211-5463.12627ArabidopsischromatinDNA methylationDRM2RdDMUAP56 |
| spellingShingle | Jacinthe Azevedo Claire Picart Laurent Dureau Dominique Pontier Sylvie Jaquinod‐Kieffer Mohamed‐Ali Hakimi Thierry Lagrange UAP56 associates with DRM2 and is localized to chromatin in Arabidopsis FEBS Open Bio Arabidopsis chromatin DNA methylation DRM2 RdDM UAP56 |
| title | UAP56 associates with DRM2 and is localized to chromatin in Arabidopsis |
| title_full | UAP56 associates with DRM2 and is localized to chromatin in Arabidopsis |
| title_fullStr | UAP56 associates with DRM2 and is localized to chromatin in Arabidopsis |
| title_full_unstemmed | UAP56 associates with DRM2 and is localized to chromatin in Arabidopsis |
| title_short | UAP56 associates with DRM2 and is localized to chromatin in Arabidopsis |
| title_sort | uap56 associates with drm2 and is localized to chromatin in arabidopsis |
| topic | Arabidopsis chromatin DNA methylation DRM2 RdDM UAP56 |
| url | https://doi.org/10.1002/2211-5463.12627 |
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