DNA damage stabilizes interaction of CSB with the transcription elongation machinery.
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.
van den Boom
Vincent
V
Department of Cell Biology and Genetics, Erasmus MC, P.O. Box 1738, 3000 DR Rotterdam, Netherlands.
Citterio
Elisabetta
E
Hoogstraten
Deborah
D
Zotter
Angelika
A
Egly
Jean-Marc
JM
van Cappellen
Wiggert A
WA
Hoeijmakers
Jan H J
JH
Houtsmuller
Adriaan B
AB
Vermeulen
Wim
W
D021581:Active Transport, Cell Nucleus; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D003057:Cockayne Syndrome / Q000378:metabolism; D003198:Computer Simulation; D004249:DNA Damage; D004265:DNA Helicases / Q000737:chemistry* / Q000378:metabolism; D004260:DNA Repair; D045643:DNA Repair Enzymes; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015151:Immunoblotting; D007700:Kinetics; D008027:Light; D008164:Luminescent Proteins / Q000378:metabolism; D008853:Microscopy; D008856:Microscopy, Fluorescence; D000075223:Poly-ADP-Ribose Binding Proteins; D011485:Protein Binding; D012319:RNA Polymerase II / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012984:Software; D013997:Time Factors; D014158:Transcription, Genetic; D014466:Ultraviolet Rays; D051760:Xeroderma Pigmentosum Group A Protein
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D008164:Luminescent Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D011993:Recombinant Fusion Proteins; C497314:XPA protein, human; D051760:Xeroderma Pigmentosum Group A Protein; D049452:Green Fluorescent Proteins; D012319:RNA Polymerase II; D004265:DNA Helicases; C078673:ERCC6 protein, human; D045643:DNA Repair Enzymes