Abstract
In the largest E3 ligase subfamily, Cul3 binds a BTB domain, and an associated protein-interaction domain such as MATH recruits substrates for ubiquitination. Here, we present biochemical and structural analyses of the MATH-BTB protein, SPOP. We define a SPOP-binding consensus (SBC) and determine structures revealing recognition of SBCs from the phosphatase Puc, the transcriptional regulator Ci, and the chromatin component MacroH2A. We identify a dimeric SPOP-Cul3 assembly involving a conserved helical structure C-terminal of BTB domains, which we call "3-box" due to its facilitating Cul3 binding and its resemblance to F-/SOCS-boxes in other cullin-based E3s. Structural flexibility between the substrate-binding MATH and Cul3-binding BTB/3-box domains potentially allows a SPOP dimer to engage multiple SBCs found within a single substrate, such as Puc. These studies provide a molecular understanding of how MATH-BTB proteins recruit substrates to Cul3 and how their dimerization and conformational variability may facilitate avid interactions with diverse substrates.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Adaptor Proteins, Signal Transducing / chemistry
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Adaptor Proteins, Signal Transducing / genetics
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Adaptor Proteins, Signal Transducing / metabolism
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Amino Acid Sequence
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Animals
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Co-Repressor Proteins
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Consensus Sequence / physiology
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Crystallography, X-Ray
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Cullin Proteins / chemistry*
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Cullin Proteins / genetics
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Cullin Proteins / metabolism
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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Drosophila Proteins / chemistry
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Drosophila Proteins / genetics
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Drosophila Proteins / metabolism
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Drosophila melanogaster
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Histones / chemistry
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Histones / genetics
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Histones / metabolism
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Humans
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Models, Molecular
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Molecular Chaperones
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Mutation / physiology
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Nuclear Proteins / chemistry*
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism
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Peptide Fragments / chemistry
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Peptide Fragments / genetics
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Peptide Fragments / metabolism
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Phosphoprotein Phosphatases / chemistry
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Phosphoprotein Phosphatases / genetics
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Phosphoprotein Phosphatases / metabolism
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Protein Binding / physiology
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Protein Interaction Domains and Motifs / physiology
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Protein Multimerization / physiology
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Protein Structure, Quaternary / physiology
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Repressor Proteins / chemistry*
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Repressor Proteins / genetics
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Repressor Proteins / metabolism
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Transcription Factors / chemistry
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Ubiquitin-Protein Ligases / chemistry*
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Ubiquitin-Protein Ligases / genetics
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Ubiquitin-Protein Ligases / metabolism
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Ubiquitination / physiology
Substances
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Adaptor Proteins, Signal Transducing
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CUL3 protein, human
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Co-Repressor Proteins
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Cullin Proteins
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DAXX protein, human
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DNA-Binding Proteins
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Drosophila Proteins
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Histones
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Molecular Chaperones
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Nuclear Proteins
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Peptide Fragments
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Recombinant Fusion Proteins
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Repressor Proteins
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SPOP protein, human
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Transcription Factors
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ci protein, Drosophila
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macroH2A histone
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Ubiquitin-Protein Ligases
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puc protein, Drosophila
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Phosphoprotein Phosphatases
Associated data
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PDB/3HQH
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PDB/3HQI
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PDB/3HQL
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PDB/3HQM
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PDB/3HSV
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PDB/3HTM
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PDB/3HU6
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PDB/3HVE
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PDB/3IVB
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PDB/3IVQ
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PDB/3IVV