Rapid switching of TFIIH between RNA polymerase I and II transcription and DNA repair in vivo.
The transcription/repair factor TFIIH operates as a DNA helix opener in RNA polymerase II (RNAP2) transcription and nucleotide excision repair. To study TFIIH in vivo, we generated cell lines expressing functional GFP-tagged TFIIH. TFIIH was homogeneously distributed throughout the nucleus with nucleolar accumulations. We provide in vivo evidence for involvement of TFIIH in RNA polymerase I (RNAP1) transcription. Photobleaching revealed that TFIIH moves freely and gets engaged in RNAP1 and RNAP2 transcription for approximately 25 and approximately 6 s, respectively. TFIIH readily switches between transcription and repair sites (where it is immobilized for approximately 4 min) without large-scale alterations in composition. Our findings support a model of diffusion and random collision of individual components that permits a quick and versatile response to changing conditions.
Hoogstraten
Deborah
D
Department of Cell Biology and Genetics, Medical Genetics Center, CBG, 3000 DR Rotterdam, The Netherlands.
Nigg
Alex L
AL
Heath
Helen
H
Mullenders
Leon H F
LH
van Driel
Roel
R
Hoeijmakers
Jan H J
JH
Vermeulen
Wim
W
Houtsmuller
Adriaan B
AB
D000818:Animals; D016466:CHO Cells; D002466:Cell Nucleolus / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D006224:Cricetinae; D003593:Cytoplasm / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D049452:Green Fluorescent Proteins; D008027:Light; D008164:Luminescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008954:Models, Biological; D012318:RNA Polymerase I / Q000737:chemistry* / Q000378:metabolism; D012319:RNA Polymerase II / Q000737:chemistry* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors; D051758:Transcription Factor TFIIH; D019473:Transcription Factors, TFII / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D014466:Ultraviolet Rays
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D019473:Transcription Factors, TFII; D049452:Green Fluorescent Proteins; D051758:Transcription Factor TFIIH; D004247:DNA; D012319:RNA Polymerase II; D012318:RNA Polymerase I