The DNA topoisomerase I binding protein topors as a novel cellular target for SUMO-1 modification: characterization of domains necessary for subcellular localization and sumolation.
Over the past years, modification by covalent attachment of SUMO (small ubiquitin-like modifier) has been demonstrated for of a number of cellular and viral proteins. While increasing evidence suggests a role for SUMO modification in the regulation of protein-protein interactions and/or subcellular localization, most SUMO targets are still at large. In this report we show that Topors, a Topoisomerase I and p53 interacting protein of hitherto unknown function, presents a novel cellular target for SUMO-1 modification. In a yeast two-hybrid system, Topors interacted with both SUMO-1 and the SUMO-1 conjugating enzyme UBC9. Multiple SUMO-1 modified forms of Topors could be detected after cotransfection of exogenous SUMO-1 and Topors induced the colocalization of a YFP tagged SUMO-1 protein in a speckled pattern in the nucleus. A subset of these Topors' nuclear speckles were closely associated with the PML nuclear bodies (POD, ND10). A central domain comprising Topors residues 437 to 574 was sufficient for both sumolation and localization to nuclear speckles. One SUMO-1 acceptor site at lysine residue 560 could be identified within this region. However, sumolation-deficient Topors mutants showed that sumolation obviously is not required for localization to nuclear speckles.
Pubmed ID: 14516784 RIS Download
Active Transport, Cell Nucleus | Bacterial Proteins | Binding Sites | Carrier Proteins | Cell Compartmentation | Cell Nucleus | Cell Nucleus Structures | DNA-Binding Proteins | Eukaryotic Cells | HeLa Cells | Humans | Ligases | Luminescent Proteins | Mutation | Neoplasm Proteins | Nuclear Proteins | Protein Structure, Tertiary | SUMO-1 Protein | Transcription Factors | Ubiquitin-Conjugating Enzymes | Ubiquitin-Protein Ligases