Mechanism of Lys48-linked polyubiquitin chain recognition by the Mud1 UBA domain.
The ubiquitin-pathway associated (UBA) domain is a 40-residue polyubiquitin-binding motif. The Schizosaccharomyces pombe protein Mud1 is an ortholog of the Saccharomyces cerevisiae DNA-damage response protein Ddi1 and binds to K48-linked polyubiquitin through its UBA domain. We have solved the cryst...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Sprog: | English |
| Udgivet: |
2005
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| _version_ | 1826290264130453504 |
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| author | Trempe, J Brown, N Lowe, E Gordon, C Campbell, I Noble, M Endicott, J |
| author_facet | Trempe, J Brown, N Lowe, E Gordon, C Campbell, I Noble, M Endicott, J |
| author_sort | Trempe, J |
| collection | OXFORD |
| description | The ubiquitin-pathway associated (UBA) domain is a 40-residue polyubiquitin-binding motif. The Schizosaccharomyces pombe protein Mud1 is an ortholog of the Saccharomyces cerevisiae DNA-damage response protein Ddi1 and binds to K48-linked polyubiquitin through its UBA domain. We have solved the crystal structure of Mud1 UBA at 1.8 angstroms resolution, revealing a canonical three-helical UBA fold. We have probed the interactions of this domain using mutagenesis, surface plasmon resonance, NMR and analytical ultracentrifugation. We show that the ubiquitin-binding surface of Mud1 UBA extends beyond previously recognized motifs and can be functionally dissected into primary and secondary ubiquitin-binding sites. Mutation of Phe330 to alanine, a residue exposed between helices 2 and 3, significantly reduces the affinity of the Mud1 UBA domain for K48-linked polyubiquitin, despite leaving the primary binding surface functionally intact. Moreover, K48-linked diubiquitin binds a single Mud1 UBA domain even in the presence of excess UBA. We therefore propose a mechanism for the recognition of K48-linked polyubiquitin chains by Mud1 in which diubiquitin units are specifically recognized by a single UBA domain. |
| first_indexed | 2024-03-07T02:41:31Z |
| format | Journal article |
| id | oxford-uuid:aa9c46f7-fa3c-4bc1-887e-39c3fc1c6766 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:41:31Z |
| publishDate | 2005 |
| record_format | dspace |
| spelling | oxford-uuid:aa9c46f7-fa3c-4bc1-887e-39c3fc1c67662022-03-27T03:16:18ZMechanism of Lys48-linked polyubiquitin chain recognition by the Mud1 UBA domain.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:aa9c46f7-fa3c-4bc1-887e-39c3fc1c6766EnglishSymplectic Elements at Oxford2005Trempe, JBrown, NLowe, EGordon, CCampbell, INoble, MEndicott, JThe ubiquitin-pathway associated (UBA) domain is a 40-residue polyubiquitin-binding motif. The Schizosaccharomyces pombe protein Mud1 is an ortholog of the Saccharomyces cerevisiae DNA-damage response protein Ddi1 and binds to K48-linked polyubiquitin through its UBA domain. We have solved the crystal structure of Mud1 UBA at 1.8 angstroms resolution, revealing a canonical three-helical UBA fold. We have probed the interactions of this domain using mutagenesis, surface plasmon resonance, NMR and analytical ultracentrifugation. We show that the ubiquitin-binding surface of Mud1 UBA extends beyond previously recognized motifs and can be functionally dissected into primary and secondary ubiquitin-binding sites. Mutation of Phe330 to alanine, a residue exposed between helices 2 and 3, significantly reduces the affinity of the Mud1 UBA domain for K48-linked polyubiquitin, despite leaving the primary binding surface functionally intact. Moreover, K48-linked diubiquitin binds a single Mud1 UBA domain even in the presence of excess UBA. We therefore propose a mechanism for the recognition of K48-linked polyubiquitin chains by Mud1 in which diubiquitin units are specifically recognized by a single UBA domain. |
| spellingShingle | Trempe, J Brown, N Lowe, E Gordon, C Campbell, I Noble, M Endicott, J Mechanism of Lys48-linked polyubiquitin chain recognition by the Mud1 UBA domain. |
| title | Mechanism of Lys48-linked polyubiquitin chain recognition by the Mud1 UBA domain. |
| title_full | Mechanism of Lys48-linked polyubiquitin chain recognition by the Mud1 UBA domain. |
| title_fullStr | Mechanism of Lys48-linked polyubiquitin chain recognition by the Mud1 UBA domain. |
| title_full_unstemmed | Mechanism of Lys48-linked polyubiquitin chain recognition by the Mud1 UBA domain. |
| title_short | Mechanism of Lys48-linked polyubiquitin chain recognition by the Mud1 UBA domain. |
| title_sort | mechanism of lys48 linked polyubiquitin chain recognition by the mud1 uba domain |
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