Abstract
The DEK oncoprotein regulates cellular chromatin function via a number of protein-protein interactions. However, the biological relevance of its unique pseudo-SAP/SAP-box domain, which transmits DNA modulating activities in vitro, remains largely speculative. As hypothesis-driven mutations failed to yield DNA-binding null (DBN) mutants, we combined random mutagenesis with the Bacterial Growth Inhibition Screen (BGIS) to overcome this bottleneck. Re-expression of a DEK-DBN mutant in newly established human DEK knockout cells failed to reduce the increase in nuclear size as compared to wild type, indicating roles for DEK-DNA interactions in cellular chromatin organization. Our results extend the functional roles of DEK in metazoan chromatin and highlight the predictive ability of recombinant protein toxicity in E. coli for unbiased studies of eukaryotic DNA modulating protein domains.
Keywords:
Escherichia coli; DEK; SAP domain; chromatin; oncogene; protein domain; protein function; recombinant protein toxicity.
© 2021 Federation of European Biochemical Societies.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Bias
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Cell Nucleus / drug effects
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Cell Size / drug effects
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Chromatin / chemistry
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Chromatin / genetics
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Chromatin / metabolism*
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Chromosomal Proteins, Non-Histone / chemistry
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Chromosomal Proteins, Non-Histone / genetics*
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Chromosomal Proteins, Non-Histone / metabolism*
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Chromosomal Proteins, Non-Histone / toxicity
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DNA / metabolism*
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Escherichia coli / drug effects*
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Escherichia coli / genetics
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Escherichia coli / growth & development
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Gene Expression Regulation, Bacterial / drug effects
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Genome, Bacterial / drug effects
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Genome, Bacterial / genetics
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Humans
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Loss of Function Mutation*
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Mutagenesis
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Nucleosomes / chemistry
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Nucleosomes / genetics
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Nucleosomes / metabolism
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Oncogene Proteins / chemistry
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Oncogene Proteins / genetics*
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Oncogene Proteins / metabolism*
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Oncogene Proteins / toxicity
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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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Peptide Fragments / toxicity
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Poly-ADP-Ribose Binding Proteins / chemistry
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Poly-ADP-Ribose Binding Proteins / genetics*
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Poly-ADP-Ribose Binding Proteins / metabolism*
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Poly-ADP-Ribose Binding Proteins / toxicity
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Protein Domains / genetics
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Recombinant Proteins / toxicity*
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Toxicity Tests / methods
Substances
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Chromatin
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Chromosomal Proteins, Non-Histone
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DEK protein, human
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Nucleosomes
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Oncogene Proteins
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Peptide Fragments
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Poly-ADP-Ribose Binding Proteins
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Recombinant Proteins
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DNA