Characterization of segments from the central region of BRCA1: an intrinsically disordered scaffold for multiple protein-protein and protein-DNA interactions?
Autor: | Blair R. Szymczyna, Jack C.C. Liao, Cheryl H. Arrowsmith, Rob C. Laister, Ying Lu, Avi Chakrabartty, Ayeda Ayed, Wen Ying Mark |
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Rok vydání: | 2004 |
Předmět: |
Protein Folding
Magnetic Resonance Spectroscopy DNA Repair Transcription Genetic DNA damage DNA repair Protein Conformation Biophysics Mutation Missense Apoptosis Breast Neoplasms Biology Polymerase Chain Reaction Biophysical Phenomena Gene product chemistry.chemical_compound Protein structure Structural Biology Humans Electrophoretic mobility shift assay Molecular Biology Dose-Response Relationship Drug Models Genetic BRCA1 Protein Circular Dichroism Cell Cycle Tryptophan DNA Protein Structure Tertiary Biochemistry chemistry Protein folding Tumor Suppressor Protein p53 Heteronuclear single quantum coherence spectroscopy Protein Binding |
Zdroj: | Journal of molecular biology. 345(2) |
ISSN: | 0022-2836 |
Popis: | The BRCA1 tumor suppressor gene encodes an 1863 amino acid gene product that is implicated in many cellular pathways including transcription, cell-cycle checkpoint control, apoptosis and DNA repair. Much attention has been focused on the structural and biochemical characterization of the N-terminal RING and tandem C-terminal BRCT domains of BRCA1. Here we used NMR spectroscopy in conjunction with CD spectroscopy and limited proteolysis to investigate the biophysical properties of the approximately 1500 residue central region of BRCA1. Our results show that although there are a few small, mildly protease-resistant regions, the majority of the BRCA1 central region lacks any pre-existing independently folded globular domains. Electrophoretic mobility shift assay and intrinsic tryptophan fluorescence experiments also demonstrate that, although intrinsically disordered, polypeptides from the central region are able to mediate interactions with DNA and p53 with affinities in the low micromolar range. This supports a model in which the central region may act as a long flexible scaffold for intermolecular interactions, thereby helping to integrate multiple signals in the DNA damage response pathway. |
Databáze: | OpenAIRE |
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