The role of lysine 369 of human topoisomerase IB in recognizing, clamping and processing its DNA substrate was experimentally investigated. Lys(369) is located in one of the two lips that interact to each other allowing the protein to embrace and firmly bind the DNA substrate. The lysine was mutated to a glutamate residue and the catalytic activity of the mutant enzyme was assayed. The mutant shows a distributive behavior, has a reduced binding to the substrate and a lower cleavage extent when compared to the wild type enzyme. The mutant displays reduced sensitivity to CPT both "in vitro" and in an "in vivo" yeast model, likely because of the low amount of cleaved DNA, however it displays cleavage and religation rates comparable to the wild type. These results demonstrate that the mutation causes a destabilization of the lips clamping around the DNA, impairing the protein-DNA interaction, emphasizing the importance of the ionic pair in tuning the stability of the protein-DNA complex.
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| Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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| Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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| Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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| Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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| Site | Modification | Modifier | Source | Reference |
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| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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| Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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| Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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| Evidence ID | Analyze ID | File | Description |
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