Structural insight into the ZFAND1-p97 interaction involved in stress granule clearance
Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved..
Arsenite-induced stress granule (SG) formation can be cleared by the ubiquitin-proteasome system aided by the ATP-dependent unfoldase p97. ZFAND1 participates in this pathway by recruiting p97 to trigger SG clearance. ZFAND1 contains two An1-type zinc finger domains (ZF1 and ZF2), followed by a ubiquitin-like domain (UBL); but their structures are not experimentally determined. To shed light on the structural basis of the ZFAND1-p97 interaction, we determined the atomic structures of the individual domains of ZFAND1 by solution-state NMR spectroscopy and X-ray crystallography. We further characterized the interaction between ZFAND1 and p97 by methyl NMR spectroscopy and cryo-EM. 15N spin relaxation dynamics analysis indicated independent domain motions for ZF1, ZF2, and UBL. The crystal structure and NMR structure of UBL showed a conserved β-grasp fold homologous to ubiquitin and other UBLs. Nevertheless, the UBL of ZFAND1 contains an additional N-terminal helix that adopts different conformations in the crystalline and solution states. ZFAND1 uses the C-terminal UBL to bind to p97, evidenced by the pronounced line-broadening of the UBL domain during the p97 titration monitored by methyl NMR spectroscopy. ZFAND1 binding induces pronounced conformational heterogeneity in the N-terminal domain of p97, leading to a partial loss of the cryo-EM density of the N-terminal domain of p97. In conclusion, this work paved the way for a better understanding of the interplay between p97 and ZFAND1 in the context of SG clearance.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:300 |
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| Enthalten in: |
The Journal of biological chemistry - 300(2024), 5 vom: 25. März, Seite 107230 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Lai, Chih-Hsuan [VerfasserIn] |
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| Links: |
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| Themen: |
AlphaFold |
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| Anmerkungen: |
Date Revised 28.04.2024 published: Print-Electronic Citation Status Publisher |
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| doi: |
10.1016/j.jbc.2024.107230 |
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NLM370272366 |
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| 520 | |a Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved. | ||
| 520 | |a Arsenite-induced stress granule (SG) formation can be cleared by the ubiquitin-proteasome system aided by the ATP-dependent unfoldase p97. ZFAND1 participates in this pathway by recruiting p97 to trigger SG clearance. ZFAND1 contains two An1-type zinc finger domains (ZF1 and ZF2), followed by a ubiquitin-like domain (UBL); but their structures are not experimentally determined. To shed light on the structural basis of the ZFAND1-p97 interaction, we determined the atomic structures of the individual domains of ZFAND1 by solution-state NMR spectroscopy and X-ray crystallography. We further characterized the interaction between ZFAND1 and p97 by methyl NMR spectroscopy and cryo-EM. 15N spin relaxation dynamics analysis indicated independent domain motions for ZF1, ZF2, and UBL. The crystal structure and NMR structure of UBL showed a conserved β-grasp fold homologous to ubiquitin and other UBLs. Nevertheless, the UBL of ZFAND1 contains an additional N-terminal helix that adopts different conformations in the crystalline and solution states. ZFAND1 uses the C-terminal UBL to bind to p97, evidenced by the pronounced line-broadening of the UBL domain during the p97 titration monitored by methyl NMR spectroscopy. ZFAND1 binding induces pronounced conformational heterogeneity in the N-terminal domain of p97, leading to a partial loss of the cryo-EM density of the N-terminal domain of p97. In conclusion, this work paved the way for a better understanding of the interplay between p97 and ZFAND1 in the context of SG clearance | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a AlphaFold | |
| 650 | 4 | |a NMR spectroscopy | |
| 650 | 4 | |a X-ray crystallography | |
| 650 | 4 | |a ZFAND1 | |
| 650 | 4 | |a chemical cross-linking | |
| 650 | 4 | |a cryo-EM | |
| 650 | 4 | |a dynamics | |
| 650 | 4 | |a p97 | |
| 650 | 4 | |a stress granule | |
| 650 | 4 | |a ubiquitin-like domain | |
| 650 | 4 | |a ubiquitin-proteasome system | |
| 650 | 4 | |a zinc finger | |
| 700 | 1 | |a Ko, Kuang-Ting |e verfasserin |4 aut | |
| 700 | 1 | |a Fan, Pei-Ju |e verfasserin |4 aut | |
| 700 | 1 | |a Yu, Tsun-Ai |e verfasserin |4 aut | |
| 700 | 1 | |a Chang, Chi-Fon |e verfasserin |4 aut | |
| 700 | 1 | |a Draczkowski, Piotr |e verfasserin |4 aut | |
| 700 | 1 | |a Hsu, Shang-Te Danny |e verfasserin |4 aut | |
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