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C/EBPα expression is partially regulated by C/EBPβ in response to DNA damage and C/EBPα-deficient fibroblasts display an impaired G1 checkpoint

Oncogene volume 28, pages 3235–3245 (2009)Cite this article

Abstract

We observed that CCAAT/enhancer-binding protein (C/EBP)α is highly inducible in primary fibroblasts by DNA-damaging agents that induce strand breaks, alkylate and crosslink DNA as well as those that produce bulky DNA lesions. Fibroblasts deficient in C/EBPα (C/EBPα−/−) display an impaired G1 checkpoint as evidenced by an inappropriate entry into the S-phase in response to DNA damage, and these cells also display an enhanced G1/S transition in response to mitogens. The induction of C/EBPα by DNA damage in fibroblasts does not require p53. Electrophoretic mobility shift assay (EMSA) analysis of nuclear extracts prepared from ultraviolet B (UVB)- and N-methyl-N′-nitro-N-nitrosoguanidine (MNNG)-treated fibroblasts showed increased binding of C/EBPβ to a C/EBP consensus sequence and chromatin immunoprecipitation (ChIP) analysis also showed increased C/EBPβ binding to the C/EBPα promoter. To determine whether C/EBPβ has a function in the regulation of C/EBPα, we treated C/EBPβ−/− fibroblasts with UVB or MNNG. We observed that C/EBPα induction was impaired in both UVB- and MNNG-treated C/EBPβ−/− fibroblasts. Our study shows a novel function for C/EBPβ in the regulation of C/EBPα in response to DNA damage and provides definitive genetic evidence that C/EBPα has a critical role in the DNA damage G1 checkpoint.

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Acknowledgements

This research was supported by grants from the National Cancer Institute (CA46637), the National Institute of Environmental Health Sciences (ES12473), and a training grant from the NIEHS (ES007046). We thank Brian Sayers for his technical support with DNA-damaging agents.

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  1. Department of Environmental and Molecular Toxicology, Cell Signaling and Cancer Group, North Carolina State University, Raleigh, NC, USA

    R Ranjan, E A Thompson & R C Smart

  2. Division of Common Cancers, National Cancer Center, Lung Cancer Branch, Goyang-si, Gyeonggi-do, South Korea,

    K Yoon

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  1. R Ranjan
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Correspondence to R C Smart.

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Ranjan, R., Thompson, E., Yoon, K. et al. C/EBPα expression is partially regulated by C/EBPβ in response to DNA damage and C/EBPα-deficient fibroblasts display an impaired G1 checkpoint. Oncogene 28, 3235–3245 (2009). https://doi.org/10.1038/onc.2009.176

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