In vivo dynamics of chromatin-associated complex formation in mammalian nucleotide excision repair

• DOI: 10.1073/pnas.0403664101
• Publication Date: 2004-11-03T02:58:58Z
• Authors: Martijn J. Moné; Tytus Bernas; Christoffel Dinant; Feliks A. Goedvree; Erik M. M. Manders; Marcel Volker; Adriaan B. Houtsm...; Jan H. J. Hoeijma...; Wim Vermeulen; Roel van Driel
• Keywords: ERCC1

Chromatin is the substrate for many processes in cell nucleus, including transcription, replication, and various DNA repair systems, all of which require formation multiprotein machineries on chromatin fiber. We have analyzed kinetics vivo assembly protein complex that responsible nucleotide excision (NER) mammalian cells. Assembly initiated by UV irradiation a small area after accumulation GFP-tagged NER proteins DNA-damaged measured, reflecting establishment dual-incision complex. The dynamic behavior two proteins, ERCC1-XPF TFIIH, was studied detail. Results show assembled with rate ≈30 complexes per second not diffusion limited. Furthermore, we provide evidence only binding but also its helicase activity, required recruitment ERCC1-XPF. These studies give quantitative insight into de novo chromatin-associated living