Identification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12
Wnt signaling is dependent on dishevelled proteins (DVL1-3), which assemble an intracellular Wnt signalosome at the plasma membrane. The levels of DVL1-3 are regulated by multiple Cullin-RING E3 ligases that mediate their ubiquitination and degradation. The BTBKelch protein KLHL12 was the first E3 u...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Language: | English |
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Royal Society
2020
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| _version_ | 1797061004981436416 |
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| author | Chen, Z Wasney, GA Picaud, S Filippakopoulos, P Vedadi, M D’Angiolella, V Bullock, AN |
| author_facet | Chen, Z Wasney, GA Picaud, S Filippakopoulos, P Vedadi, M D’Angiolella, V Bullock, AN |
| author_sort | Chen, Z |
| collection | OXFORD |
| description | Wnt signaling is dependent on dishevelled proteins (DVL1-3), which assemble an
intracellular Wnt signalosome at the plasma membrane. The levels of DVL1-3 are regulated by
multiple Cullin-RING E3 ligases that mediate their ubiquitination and degradation. The BTBKelch protein KLHL12 was the first E3 ubiquitin ligase to be identified for DVL1-3, but the
molecular mechanisms determining its substrate interactions have remained unknown. Here, we
mapped the interaction of DVL1-3 to a ‘PGXPP’ motif that is conserved in other known partners
and substrates of KLHL12, including PLEKHA4, PEF1, SEC31 and DRD4. To determine the
binding mechanism, we solved a 2.4 Å crystal structure of the Kelch domain of KLHL12 in
complex with a DVL1 peptide that bound with low micromolar affinity. The DVL1 substrate
adopted a U-shaped turn conformation that enabled hydrophobic interactions with all six blades
of the Kelch domain β-propeller. In cells, the mutation or deletion of this motif reduced the
binding and ubiquitination of DVL1 and increased its stability confirming this sequence as a
degron motif for KLHL12 recruitment. These results define the molecular mechanisms
determining DVL regulation by KLHL12 and establish the KLHL12 Kelch domain as a new
protein interaction module for a novel proline-rich motif. |
| first_indexed | 2024-03-06T20:24:55Z |
| format | Journal article |
| id | oxford-uuid:2f13113b-5f94-44f0-9b1e-2e341b4fe111 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T20:24:55Z |
| publishDate | 2020 |
| publisher | Royal Society |
| record_format | dspace |
| spelling | oxford-uuid:2f13113b-5f94-44f0-9b1e-2e341b4fe1112022-03-26T12:53:01ZIdentification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:2f13113b-5f94-44f0-9b1e-2e341b4fe111EnglishSymplectic ElementsRoyal Society 2020Chen, ZWasney, GAPicaud, SFilippakopoulos, PVedadi, MD’Angiolella, VBullock, ANWnt signaling is dependent on dishevelled proteins (DVL1-3), which assemble an intracellular Wnt signalosome at the plasma membrane. The levels of DVL1-3 are regulated by multiple Cullin-RING E3 ligases that mediate their ubiquitination and degradation. The BTBKelch protein KLHL12 was the first E3 ubiquitin ligase to be identified for DVL1-3, but the molecular mechanisms determining its substrate interactions have remained unknown. Here, we mapped the interaction of DVL1-3 to a ‘PGXPP’ motif that is conserved in other known partners and substrates of KLHL12, including PLEKHA4, PEF1, SEC31 and DRD4. To determine the binding mechanism, we solved a 2.4 Å crystal structure of the Kelch domain of KLHL12 in complex with a DVL1 peptide that bound with low micromolar affinity. The DVL1 substrate adopted a U-shaped turn conformation that enabled hydrophobic interactions with all six blades of the Kelch domain β-propeller. In cells, the mutation or deletion of this motif reduced the binding and ubiquitination of DVL1 and increased its stability confirming this sequence as a degron motif for KLHL12 recruitment. These results define the molecular mechanisms determining DVL regulation by KLHL12 and establish the KLHL12 Kelch domain as a new protein interaction module for a novel proline-rich motif. |
| spellingShingle | Chen, Z Wasney, GA Picaud, S Filippakopoulos, P Vedadi, M D’Angiolella, V Bullock, AN Identification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12 |
| title | Identification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12 |
| title_full | Identification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12 |
| title_fullStr | Identification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12 |
| title_full_unstemmed | Identification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12 |
| title_short | Identification of a PGXPP degron motif in dishevelled and structural basis for its binding to the E3 ligase KLHL12 |
| title_sort | identification of a pgxpp degron motif in dishevelled and structural basis for its binding to the e3 ligase klhl12 |
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