Recruitment of phosphorylated chromatin assembly factor 1 to chromatin after UV irradiation of human cells.
The subcellular distribution and posttranslational modification of human chromatin assembly factor 1 (CAF-1) have been investigated after UV irradiation of HeLa cells. In an asynchronous cell population only a subfraction of the two large CAF-1 subunits, p150 and p60, were found to exist in a chromatin-associated fraction. This fraction is most abundant during S phase in nonirradiated cells and is much reduced in G2 cells. After UV irradiation, the chromatin-associated form of CAF-1 dramatically increased in all cells irrespective of their position in the cell cycle. Such chromatin recruitment resembles that seen for PCNA, a DNA replication and repair factor. The chromatin-associated fraction of p60 was predominantly hypophosphorylated in nonirradiated G2 cells. UV irradiation resulted in the rapid recruitment to chromatin of phosphorylated forms of the p60 subunit. Furthermore, the amount of the p60 and p150 subunits of CAF-1 associated with chromatin was a function of the dose of UV irradiation. Consistent with these in vivo observations, we found that the amount of CAF-1 required to stimulate nucleosome assembly during the repair of UV photoproducts in vitro depended upon both the number of lesions and the phosphorylation state of CAF-1. The recruitment of CAF-1 to chromatin in response to UV irradiation of human cells described here supports a physiological role for CAF-1 in linking chromatin assembly to DNA repair.
Martini
E
E
Institut Curie, Research Section Unité Mixte de Recherche, 144 du Centre National de la Recherche Scientifique, 75231 Paris Cedex 05, France.
Roche
D M
DM
Marheineke
K
K
Verreault
A
A
Almouzni
G
G
D000818:Animals; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism* / Q000528:radiation effects; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins / Q000378:metabolism* / Q000502:physiology / Q000528:radiation effects; D016195:G2 Phase; D006367:HeLa Cells; D006801:Humans; D017830:Octoxynol; D010766:Phosphorylation; D018809:Proliferating Cell Nuclear Antigen / Q000502:physiology; D011817:Rabbits; D014157:Transcription Factors; D014466:Ultraviolet Rays
C533593:CHAF1B protein, human; C489780:CNOT8 protein, human; D002843:Chromatin; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D018809:Proliferating Cell Nuclear Antigen; D014157:Transcription Factors; D017830:Octoxynol