Oligomerization mediated by the D2 domain of DTX3L is critical for DTX3L-PARP9 reading function of mono-ADP-ribosylated androgen receptor
© 2024 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society..
Deltex proteins are a family of E3 ubiquitin ligases that encode C-terminal RING and DTC domains that mediate interactions with E2 ubiquitin-conjugating enzymes and recognize ubiquitination substrates. DTX3L is unique among the Deltex proteins based on its N-terminal domain architecture. The N-terminal D1 and D2 domains of DTX3L mediate homo-oligomerization, and the D3 domain interacts with PARP9, a protein that contains tandem macrodomains with ADP-ribose reader function. While DTX3L and PARP9 are known to heterodimerize, and assemble into a high molecular weight oligomeric complex, the nature of the oligomeric structure, including whether this contributes to the ADP-ribose reader function is unknown. Here, we report a crystal structure of the DTX3L N-terminal D2 domain and show that it forms a tetramer with, conveniently, D2 symmetry. We identified two interfaces in the structure: a major, conserved interface with a surface of 973 Å2 and a smaller one of 415 Å2. Using native mass spectrometry, we observed molecular species that correspond to monomers, dimers and tetramers of the D2 domain. Reconstitution of DTX3L knockout cells with a D1-D2 deletion mutant showed the domain is dispensable for DTX3L-PARP9 heterodimer formation, but necessary to assemble an oligomeric complex with efficient reader function for ADP-ribosylated androgen receptor. Our results suggest that homo-oligomerization of DTX3L is important for the DTX3L-PARP9 complex to read mono-ADP-ribosylation on a ligand-regulated transcription factor.
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| Medienart: |
E-Artikel |
| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:33 |
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| Enthalten in: |
Protein science : a publication of the Protein Society - 33(2024), 4 vom: 02. Apr., Seite e4945 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Vela-Rodríguez, Carlos [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 22.03.2024 Date Revised 01.04.2024 published: Print UpdateOf: bioRxiv. 2023 Nov 29;:. - PMID 38076829 Citation Status MEDLINE |
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| doi: |
10.1002/pro.4945 |
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| 100 | 1 | |a Vela-Rodríguez, Carlos |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Oligomerization mediated by the D2 domain of DTX3L is critical for DTX3L-PARP9 reading function of mono-ADP-ribosylated androgen receptor |
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| 500 | |a UpdateOf: bioRxiv. 2023 Nov 29;:. - PMID 38076829 | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2024 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society. | ||
| 520 | |a Deltex proteins are a family of E3 ubiquitin ligases that encode C-terminal RING and DTC domains that mediate interactions with E2 ubiquitin-conjugating enzymes and recognize ubiquitination substrates. DTX3L is unique among the Deltex proteins based on its N-terminal domain architecture. The N-terminal D1 and D2 domains of DTX3L mediate homo-oligomerization, and the D3 domain interacts with PARP9, a protein that contains tandem macrodomains with ADP-ribose reader function. While DTX3L and PARP9 are known to heterodimerize, and assemble into a high molecular weight oligomeric complex, the nature of the oligomeric structure, including whether this contributes to the ADP-ribose reader function is unknown. Here, we report a crystal structure of the DTX3L N-terminal D2 domain and show that it forms a tetramer with, conveniently, D2 symmetry. We identified two interfaces in the structure: a major, conserved interface with a surface of 973 Å2 and a smaller one of 415 Å2. Using native mass spectrometry, we observed molecular species that correspond to monomers, dimers and tetramers of the D2 domain. Reconstitution of DTX3L knockout cells with a D1-D2 deletion mutant showed the domain is dispensable for DTX3L-PARP9 heterodimer formation, but necessary to assemble an oligomeric complex with efficient reader function for ADP-ribosylated androgen receptor. Our results suggest that homo-oligomerization of DTX3L is important for the DTX3L-PARP9 complex to read mono-ADP-ribosylation on a ligand-regulated transcription factor | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a ADP‐ribosylation | |
| 650 | 4 | |a DTX3L | |
| 650 | 4 | |a androgen receptor | |
| 650 | 4 | |a crystal structure | |
| 650 | 4 | |a oligomerization | |
| 650 | 4 | |a ubiquitination | |
| 650 | 7 | |a Receptors, Androgen |2 NLM | |
| 650 | 7 | |a Ubiquitin-Protein Ligases |2 NLM | |
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| 650 | 7 | |a Adenosine Diphosphate Ribose |2 NLM | |
| 650 | 7 | |a 20762-30-5 |2 NLM | |
| 700 | 1 | |a Yang, Chunsong |e verfasserin |4 aut | |
| 700 | 1 | |a Alanen, Heli I |e verfasserin |4 aut | |
| 700 | 1 | |a Eki, Rebeka |e verfasserin |4 aut | |
| 700 | 1 | |a Abbas, Tarek A |e verfasserin |4 aut | |
| 700 | 1 | |a Maksimainen, Mirko M |e verfasserin |4 aut | |
| 700 | 1 | |a Glumoff, Tuomo |e verfasserin |4 aut | |
| 700 | 1 | |a Duman, Ramona |e verfasserin |4 aut | |
| 700 | 1 | |a Wagner, Armin |e verfasserin |4 aut | |
| 700 | 1 | |a Paschal, Bryce M |e verfasserin |4 aut | |
| 700 | 1 | |a Lehtiö, Lari |e verfasserin |4 aut | |
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