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.

[1]  K. Marheineke,et al.  Nucleosome Assembly Activity and Intracellular Localization of Human CAF-1 Changes during the Cell Division Cycle* , 1998, The Journal of Biological Chemistry.

[2]  J. Berman,et al.  Chromatin assembly factor I contributes to the maintenance, but not the re-establishment, of silencing at the yeast silent mating loci. , 1998, Genes & development.

[3]  Bruce Stillman,et al.  Nucleosomal DNA regulates the core-histone-binding subunit of the human Hat1 acetyltransferase , 1998, Current Biology.

[4]  D. S. Henderson,et al.  Chromosome fragmentation resulting from an inability to repair transposase-induced DNA double-strand breaks in PCNA mutants of Drosophila. , 1998, Mutagenesis.

[5]  P. Nurse.  Checkpoint Pathways Come of Age , 1997, Cell.

[6]  L. Cox.  Who binds wins: Competition for PCNA rings out cell-cycle changes. , 1997, Trends in cell biology.

[7]  D. de Bruin,et al.  The yeast Cac1 protein is required for the stable inheritance of transcriptionally repressed chromatin at telomeres. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[8]  R. Wood,et al.  Initiation and bidirectional propagation of chromatin assembly from a target site for nucleotide excision repair , 1997, The EMBO journal.

[9]  Richard D. Wood,et al.  Nucleotide Excision Repair in Mammalian Cells* , 1997, The Journal of Biological Chemistry.

[10]  T. Richmond,et al.  Crystal structure of the nucleosome core particle at 2.8 Å resolution , 1997, Nature.

[11]  W. Gehring,et al.  The cramped gene of Drosophila is a member of the Polycomb-group, and interacts with mus209, the gene encoding Proliferating Cell Nuclear Antigen. , 1997, Development.

[12]  Matthias Mann,et al.  Chromatin-remodelling factor CHRAC contains the ATPases ISWI and topoisomerase II , 1997, Nature.

[13]  V. Pirrotta,et al.  Chromatin-silencing mechanisms in Drosophila maintain patterns of gene expression. , 1997, Trends in genetics : TIG.

[14]  Ryuji Kobayashi,et al.  ACF, an ISWI-Containing and ATP-Utilizing Chromatin Assembly and Remodeling Factor , 1997, Cell.

[15]  T. Tsukiyama,et al.  Chromatin remodeling and transcription. , 1997, Current opinion in genetics & development.

[16]  R. Kobayashi,et al.  Ultraviolet radiation sensitivity and reduction of telomeric silencing in Saccharomyces cerevisiae cells lacking chromatin assembly factor-I. , 1997, Genes & development.

[17]  J. Berman,et al.  RLF2, a subunit of yeast chromatin assembly factor-I, is required for telomeric chromatin function in vivo. , 1997, Genes & development.

[18]  G. Pfeifer.  Formation and Processing of UV Photoproducts: Effects of DNA Sequence and Chromatin Environment , 1997, Photochemistry and photobiology.

[19]  J. Pines,et al.  Cyclin/Cdk-Dependent Initiation of DNA Replication in a Human Cell-Free System , 1997, Cell.

[20]  R. Kobayashi,et al.  Nucleosome Assembly by a Complex of CAF-1 and Acetylated Histones H3/H4 , 1996, Cell.

[21]  C. Allis,et al.  Histone Acetylation and Chromatin Assembly: A Single Escort, Multiple Dances? , 1996, Cell.

[22]  B. Stillman,et al.  Chromatin Assembly Coupled to DNA Repair: A New Role for Chromatin Assembly Factor I , 1996, Cell.

[23]  P. Kaufman.  Nucleosome assembly: the CAF and the HAT. , 1996, Current opinion in cell biology.

[24]  D. Lane,et al.  Two Pathways for Base Excision Repair in Mammalian Cells (*) , 1996, The Journal of Biological Chemistry.

[25]  B. Stillman,et al.  Postreplicative chromatin assembly by Drosophila and human chromatin assembly factor 1 , 1996, Molecular and cellular biology.

[26]  R. Wood,et al.  Detection of nucleotide excision repair incisions in human fibroblasts by immunostaining for PCNA. , 1995, Experimental cell research.

[27]  T. Krude.  Chromatin: Nucleosome assembly during DNA replication , 1995, Current Biology.

[28]  T. Krude.  Chromatin assembly factor 1 (CAF-1) colocalizes with replication foci in HeLa cell nuclei. , 1995, Experimental cell research.

[29]  P. Burgers,et al.  A mutational analysis of the yeast proliferating cell nuclear antigen indicates distinct roles in DNA replication and DNA repair , 1995, Molecular and cellular biology.

[30]  Bruce Stillman,et al.  The p150 and p60 subunits of chromatin assemblyfactor I: A molecular link between newly synthesized histories and DNA replication , 1995, Cell.

[31]  A. Attaran,et al.  BM28, a human member of the MCM2-3-5 family, is displaced from chromatin during DNA replication , 1995, The Journal of cell biology.

[32]  R. Wood,et al.  Detection and measurement of nucleotide excision repair synthesis by mammalian cell extracts in vitro , 1995 .

[33]  A. Sancar.  DNA repair in humans. , 1995, Annual review of genetics.

[34]  A. Goldman,et al.  Meiotic recombination hotspots. , 1995, Annual review of genetics.

[35]  M. P. Carty,et al.  UV light‐induced DNA synthesis arrest in HeLa cells is associated with changes in phosphorylation of human single‐stranded DNA‐binding protein. , 1994, The EMBO journal.

[36]  D. S. Henderson,et al.  Mutagen sensitivity and suppression of position‐effect variegation result from mutations in mus209, the Drosophila gene encoding PCNA. , 1994, The EMBO journal.

[37]  P. Herrlich,et al.  The mammalian UV response: mechanism of DNA damage induced gene expression. , 1994, Advances in enzyme regulation.

[38]  V. F. Liu,et al.  The ionizing radiation-induced replication protein A phosphorylation response differs between ataxia telangiectasia and normal human cells , 1993, Molecular and cellular biology.

[39]  L. Stivala,et al.  Proliferating cell nuclear antigen complex formation induced by ultraviolet irradiation in human quiescent fibroblasts as detected by immunostaining and flow cytometry. , 1993, Experimental cell research.

[40]  Y. Takasaki,et al.  Two types of proliferating cell nuclear antigen (PCNA) complex formation in quiescent normal and xeroderma pigmentosum group A fibroblasts following ultraviolet light (uv) irradiation. , 1992, Experimental cell research.

[41]  D L Spector,et al.  Dynamic organization of DNA replication in mammalian cell nuclei: spatially and temporally defined replication of chromosome-specific alpha-satellite DNA sequences , 1992, The Journal of cell biology.

[42]  Oscar M. Aparicio,et al.  Modifiers of position effect are shared between telomeric and silent mating-type loci in S. cerevisiae , 1991, Cell.

[43]  B. Stillman,et al.  Immunological characterization of chromatin assembly factor I, a human cell factor required for chromatin assembly during DNA replication in vitro. , 1991, The Journal of biological chemistry.

[44]  M. Smerdon,et al.  DNA repair and the role of chromatin structure. , 1991, Current opinion in cell biology.

[45]  B. Stillman,et al.  Purification and characterization of CAF-I, a human cell factor required for chromatin assembly during DNA replication in vitro , 1989, Cell.

[46]  R. Bravo,et al.  Changes in cyclin/proliferating cell nuclear antigen distribution during DNA repair synthesis , 1988, The Journal of cell biology.

[47]  R. Bravo,et al.  Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites , 1987, The Journal of cell biology.

[48]  B. Stillman.  Chromatin assembly during SV40 DNA replication in vitro , 1986, Cell.

[49]  S. Penman,et al.  The nonchromatin substructures of the nucleus: the ribonucleoprotein (RNP)-containing and RNP-depleted matrices analyzed by sequential fractionation and resinless section electron microscopy , 1986, The Journal of cell biology.

[50]  J. Li,et al.  Simian virus 40 DNA replication in vitro. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[51]  D. Pettijohn,et al.  Human-specific nuclear protein that associates with the polar region of the mitotic apparatus: Distribution in a human/hamster hybrid cell , 1980, Cell.

[52]  W. Earnshaw,et al.  Nucleosome assembly , 1980, Nature.

[53]  B. Hamkalo,et al.  Chromatin Structure and Function , 1979, NATO Advanced Study Institutes Series.

[54]  D. Brutlag,et al.  Nicking-closing enzyme assembles nucleosome-like structures in vitro. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[55]  M. Smerdon,et al.  Nucleosome rearrangement in human chromatin during UV-induced DNA- reapir synthesis. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[56]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.