DNA damage stabilizes interaction of CSB with the transcription elongation machinery
January 2004
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The Cockayne syndrome B (CSB) protein is essential for transcription-coupled DNA repair (TCR), which is dependent on RNA polymerase II elongation. TCR is required to quickly remove the cytotoxic transcription-blocking DNA lesions. Functional GFP-tagged CSB, expressed at physiological levels, was homogeneously dispersed throughout the nucleoplasm in addition to bright nuclear foci and nucleolar accumulation. Photobleaching studies showed that GFP-CSB, as part of a high molecular weight complex, transiently interacts with the transcription machinery. Upon (DNA damage-induced) transcription arrest CSB binding these interactions are prolonged, most likely reflecting actual engagement of CSB in TCR. These findings are consistent with a model in which CSB monitors progression of transcription by regularly probing elongation complexes and becomes more tightly associated to these complexes when TCR is active.
- Light
- Cell Line
- Humans
- Research Support, Non-U.S. Gov't
- Green Fluorescent Proteins
- Protein Binding
- Xeroderma Pigmentosum Group A Protein
- Ultraviolet Rays
- Time Factors
- *Transcription, Genetic
- Cell Nucleus/metabolism
- Microscopy, Fluorescence
- DNA-Binding Proteins/genetics
- Cells, Cultured
- DNA Repair
- Immunoblotting
- Kinetics
- Fibroblasts/metabolism
- Active Transport, Cell Nucleus
- *DNA Damage
- Image Processing, Computer-Assisted
- Computer Simulation
- DNA, Complementary/metabolism
- Luminescent Proteins/metabolism
- Cockayne Syndrome/metabolism
- DNA Helicases/*chemistry/metabolism
- Microscopy
- RNA Polymerase II/chemistry
- Recombinant Fusion Proteins/chemistry/metabolism
- Software
- transcription
- gfp-csb
- protein
- repair
- rnap ii
- polymerase
- fraction
- immobilization
- molecule
- elongation
- factor
- rna polymerase ii
- mobility
- lesion
- analysis
- uv irradiation
- complex
- damage
- syndrome
- cockayne
