Identification of RNF114 as ADPr-Ub reader through non-hydrolysable ubiquitinated ADP-ribose
Abstract Crosstalk between the post-translational modification processes of ubiquitination and ADP-ribosylation occurs in DNA-damage- and immune-responses, in addition the physical linkage of ADP-ribose and ubiquitin is found during bacterial infection. Here, we study the ubiquitination of ADP-ribos...
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| Format: | Article |
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Nature Portfolio
2025-07-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-61111-7 |
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| author | Max S. Kloet Chatrin Chatrin Rishov Mukhopadhyay Bianca D. M. van Tol Rebecca Smith Sarah A. Rotman Rayman T. N. Tjokrodirijo Kang Zhu Andrii Gorelik Lucy Maginn Paul R. Elliott Peter A. van Veelen Dragana Ahel Ivan Ahel Gerbrand J. van der Heden van Noort |
| author_facet | Max S. Kloet Chatrin Chatrin Rishov Mukhopadhyay Bianca D. M. van Tol Rebecca Smith Sarah A. Rotman Rayman T. N. Tjokrodirijo Kang Zhu Andrii Gorelik Lucy Maginn Paul R. Elliott Peter A. van Veelen Dragana Ahel Ivan Ahel Gerbrand J. van der Heden van Noort |
| author_sort | Max S. Kloet |
| collection | DOAJ |
| description | Abstract Crosstalk between the post-translational modification processes of ubiquitination and ADP-ribosylation occurs in DNA-damage- and immune-responses, in addition the physical linkage of ADP-ribose and ubiquitin is found during bacterial infection. Here, we study the ubiquitination of ADP-ribose mediated by human Deltex E3 ligases and the subsequent fate of the formed hybrid post-translational modification. We prepare a non-hydrolysable ADPr-Ub probe that we employ in a proteomics approach and identify RNF114 as an interacting protein. Using biophysical and biochemical experiments, we validate that RNF114 preferentially interacts with ubiquitinated ADP-ribose over non-modified ubiquitin. Subsequently, RNF114 can elongate the ubiquitinated ADP-ribose with a K11-linked ubiquitin chain. Using domain deletion analysis, we pinpoint the tandem zinc fingers and ubiquitin interacting motif (ZnF2 + ZnF3+UIM) domains of RNF114 to be crucial for recognising ubiquitinated ADP-ribose. Moreover, these domains are essential for the recruitment of RNF114 to the sites of laser-induced DNA damage. |
| format | Article |
| id | doaj-art-cdda0832a0ba4b2ea26ec81d4cf6898f |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-cdda0832a0ba4b2ea26ec81d4cf6898f2025-08-20T04:02:55ZengNature PortfolioNature Communications2041-17232025-07-0116111410.1038/s41467-025-61111-7Identification of RNF114 as ADPr-Ub reader through non-hydrolysable ubiquitinated ADP-riboseMax S. Kloet0Chatrin Chatrin1Rishov Mukhopadhyay2Bianca D. M. van Tol3Rebecca Smith4Sarah A. Rotman5Rayman T. N. Tjokrodirijo6Kang Zhu7Andrii Gorelik8Lucy Maginn9Paul R. Elliott10Peter A. van Veelen11Dragana Ahel12Ivan Ahel13Gerbrand J. van der Heden van Noort14Department of Cell and Chemical Biology, Leiden University Medical CentreSir William Dunn School of Pathology, University of OxfordDepartment of Cell and Chemical Biology, Leiden University Medical CentreDepartment of Cell and Chemical Biology, Leiden University Medical CentreSir William Dunn School of Pathology, University of OxfordCentre for Proteomics and Metabolomics, Leiden University Medical CentreCentre for Proteomics and Metabolomics, Leiden University Medical CentreSir William Dunn School of Pathology, University of OxfordSir William Dunn School of Pathology, University of OxfordSir William Dunn School of Pathology, University of OxfordDepartment of Biochemistry, University of OxfordCentre for Proteomics and Metabolomics, Leiden University Medical CentreSir William Dunn School of Pathology, University of OxfordSir William Dunn School of Pathology, University of OxfordDepartment of Cell and Chemical Biology, Leiden University Medical CentreAbstract Crosstalk between the post-translational modification processes of ubiquitination and ADP-ribosylation occurs in DNA-damage- and immune-responses, in addition the physical linkage of ADP-ribose and ubiquitin is found during bacterial infection. Here, we study the ubiquitination of ADP-ribose mediated by human Deltex E3 ligases and the subsequent fate of the formed hybrid post-translational modification. We prepare a non-hydrolysable ADPr-Ub probe that we employ in a proteomics approach and identify RNF114 as an interacting protein. Using biophysical and biochemical experiments, we validate that RNF114 preferentially interacts with ubiquitinated ADP-ribose over non-modified ubiquitin. Subsequently, RNF114 can elongate the ubiquitinated ADP-ribose with a K11-linked ubiquitin chain. Using domain deletion analysis, we pinpoint the tandem zinc fingers and ubiquitin interacting motif (ZnF2 + ZnF3+UIM) domains of RNF114 to be crucial for recognising ubiquitinated ADP-ribose. Moreover, these domains are essential for the recruitment of RNF114 to the sites of laser-induced DNA damage.https://doi.org/10.1038/s41467-025-61111-7 |
| spellingShingle | Max S. Kloet Chatrin Chatrin Rishov Mukhopadhyay Bianca D. M. van Tol Rebecca Smith Sarah A. Rotman Rayman T. N. Tjokrodirijo Kang Zhu Andrii Gorelik Lucy Maginn Paul R. Elliott Peter A. van Veelen Dragana Ahel Ivan Ahel Gerbrand J. van der Heden van Noort Identification of RNF114 as ADPr-Ub reader through non-hydrolysable ubiquitinated ADP-ribose Nature Communications |
| title | Identification of RNF114 as ADPr-Ub reader through non-hydrolysable ubiquitinated ADP-ribose |
| title_full | Identification of RNF114 as ADPr-Ub reader through non-hydrolysable ubiquitinated ADP-ribose |
| title_fullStr | Identification of RNF114 as ADPr-Ub reader through non-hydrolysable ubiquitinated ADP-ribose |
| title_full_unstemmed | Identification of RNF114 as ADPr-Ub reader through non-hydrolysable ubiquitinated ADP-ribose |
| title_short | Identification of RNF114 as ADPr-Ub reader through non-hydrolysable ubiquitinated ADP-ribose |
| title_sort | identification of rnf114 as adpr ub reader through non hydrolysable ubiquitinated adp ribose |
| url | https://doi.org/10.1038/s41467-025-61111-7 |
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