Nucleic Acids Research, Vol 27, Issue 19 3881-3890, Copyright © 1999 by Oxford University Press
DM Graunke, AJ Fornace Jr and RO Pieper
GADD45 has been suggested to coordinate cell cycle regulation with the
repair of DNA damage following ionizing radiation (IR). Although the GADD45
gene is transcriptionally up-regulated in response to IR, alterations in in
vivo transcription factor (TF) binding or chromatin structure associated
with up-regulation have not been defined. To understand how chromatin
structure might influence TF binding and GADD45 up-regulation, key
regulatory regions of the gene were identified by in vivo DNase I
hypersensitivity (HS) analysis. Chromatin structure and in vivo TF binding
in these regions were subsequently monitored in both non-irradiated and
irradiated human ML-1 cells. In non-irradiated cells expressing basal
levels of GADD45, the gene exhibited a highly organized chromatin structure
with distinctly positioned nucleosomes. Also identified in non-irradiated
cells were DNA-protein interactions at octamer binding motifs and a CCAAT
box in the promoter and at consensus binding sites for AP-1 and p53 within
intron 3.Upon irradiation and a subsequent 15-fold increase in GADD45 mRNA
levels, neither the chromatin structure nor the pattern of TF binding in
key regulatory regions was altered. These results suggest that the GADD45
gene is poised for up-regulation and can be rapidly induced independent of
gross changes in chromatin structure or TF binding.
ARTICLES
Presetting of chromatin structure and transcription factor binding poise the human GADD45 gene for rapid transcriptional up-regulation
DNA Damage and Repair, Pennington Biomedical Research Center, Louisiana State University, 6400 Perkins Road, Room C2058-7, Baton Rouge, LA 70808, USA.
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