p53 responsive nucleotide excision repair gene products p48 and XPC, but not p53, localize to sites of UV-irradiation-induced DNA damage, in vivo

Fitch, Maureen E.; Cross, Irina V.; Ford, James M.

doi:10.1093/carcin/bgg031

Cited by 73 publications

(45 citation statements)

References 49 publications

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“…This is not surprising as the underlying mechanism to initiate productive repair is based on the binding of the heterodimeric protein to DNA damage. Earlier studies have reported that DDB2 is required for the recruitment of exogenously transfected DDB1 to the local UV-induced damage sites (33). Here, we tested the chromatin binding of endogenous DDB1 protein following UV irradiation and show that almost all the native DDB1 in the nuclei translocates from loosely bound to a tightly bound chromatin form upon UV irradiation.…”

Section: Discussionmentioning

confidence: 83%

“…Previous studies have shown that DDB1 is not needed for the repair of damage in the NER system reconstituted in vitro (19,20). However, in vivo studies have shown that ectopically expressed DDB1 could be recruited to the damage sites following UV irradiation (33), suggesting that DDB1, as a subunit of DDB, is capable of binding to the damaged sites. In this study, we investigated the chromatin binding and UV-induced translocation of DDB1 by using cellular fractionation and immunoblot analysis.…”

Section: Resultsmentioning

confidence: 98%

“…It is already known that DDB1, DDB2, and Cul4A form a complex with an inherent E3 ubiquitin ligase activity (25), and three subunits of this complex can be recruited to UV-induced damage sites (14,33,36). Nevertheless, the role of DDB1 in the UV-induced recruitment of Cul4A and/or DDB2 is still unclear.…”

Section: Resultsmentioning

confidence: 99%

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DNA Damage Binding Protein Component DDB1 Participates in Nucleotide Excision Repair through DDB2 DNA-binding and Cullin 4A Ubiquitin Ligase Activity

Wang²,

Zhu³

et al. 2006

Cancer Research

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Functional defect in DNA damage binding (DDB) activity has a direct relationship to decreased nucleotide excision repair (NER) and increased susceptibility to cancer. DDB forms a complex with cullin 4A (Cul4A), which is now known to ubiquitylate DDB2, XPC, and histone H2A. However, the exact role of DDB1 in NER is unclear. In this study, we show that DDB1 knockdown in human cells impaired their ability to efficiently repair UV-induced cyclobutane pyrimidine dimers (CPD) but not 6-4 photoproducts (6-4PP). Extensive nuclear protein fractionation and chromatin association analysis revealed that upon irradiation, DDB1 protein is translocated from a loosely bound to a tightly bound in vivo chromatin fraction and the DDB1 translocation required the participation of functional DDB2 protein. DDB1 knockdown also affected the translocation of Cul4A component to the tightly bound form in UV-damaged chromatin in vivo as well as its recruitment to the locally damaged nuclear foci in situ. However, DDB1 knockdown had no effect on DNA damage binding capacity of DDB2. The data indicated that DDB2 can bind to damaged DNA in vivo as a monomer, whereas Cul4A recruitment to damage sites depends on the fully assembled complex. Our data also showed that DDB1 is required for the UV-induced DDB2 ubiquitylation and degradation. In summary, the results suggest that (a) DDB1 is critical for efficient NER of CPD; (b) DDB1 acts in bridging DDB2 and ubiquitin ligase Cul4A; and (c) DDB1 aids in recruiting the ubiquitin ligase activity to the damaged sites for successful commencement of lesion processing by NER. (Cancer Res 2006; 66(17): 8590-7)

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Section: Discussionmentioning

confidence: 83%

Section: Resultsmentioning

confidence: 98%

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DNA Damage Binding Protein Component DDB1 Participates in Nucleotide Excision Repair through DDB2 DNA-binding and Cullin 4A Ubiquitin Ligase Activity

Wang²,

Zhu³

et al. 2006

Cancer Research

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“…Using local irradiation, an elegant technique that allows visualization of events occurring in UVC-irradiated zones of the nuclei (Moné et al, 2001), we show that PARP activation occurs at the site of UVC exposure as rapidly as reported accumulation of p48/UV DNA damage binding protein 2 (UV-DDB2) (Fitch et al, 2003;Tang and Chu, 2002). Since DDB protein complex is thought to be responsible for initial recognition of CPD damage (Hanawalt, 2002;Tang and Chu, 2002), our results indicate that PARP Journal of Cell Science 118 (3) activation occurs concomitant with DDB2 binding to the direct DNA lesions and occurs prior to the start of NER process at the damaged site.…”

Section: Discussionmentioning

confidence: 99%

Mechanism of early biphasic activation of poly(ADP-ribose) polymerase-1 in response to ultraviolet B radiation

Vodenicharov

Ghodgaonkar

Halappanavar

et al. 2005

Journal of Cell Science

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The damage to DNA caused by ultraviolet B radiation (280-320 nm) contributes significantly to development of sunlight-induced skin cancers. The susceptibility of mice to ultraviolet B-induced skin carcinogenesis is increased by an inhibitor of the DNA damage-activated nuclear enzyme poly(ADP-ribose) polymerase-1 (PARP), hence PARP activation is likely to be associated with cellular responses that suppress carcinogenesis. To understand the role of activated PARP in these cellular functions, we need to first clearly identify the cause of PARP activation in ultraviolet B-irradiated cells. Ultraviolet B, like ultraviolet C, causes direct DNA damage of cyclobutane pyrimidine dimer and 6, 4-photoproduct types, which are subjected to the nucleotide excision repair. Moreover, ultraviolet B also causes oxidative DNA damage, which is subjected to base excision repair. To identify which of these two types of DNA damage activates PARP, we examined mechanism of early PARP activation in mouse fibroblasts exposed to ultraviolet B and C radiations. The ultraviolet B-irradiated cells rapidly activated PARP in two distinct phases, initially within the first 5 minutes and later between 60-120 minutes, whereas ultraviolet C-irradiated cells showed only the immediate PARP activation. Using antioxidants, local irradiation, chromatin immunoprecipitation and in vitro PARP assays, we identified that ultraviolet radiation-induced direct DNA damage, such as thymine dimers, cause the initial PARP activation, whereas ultraviolet B-induced oxidative damage cause the second PARP activation. Our results suggest that cells can selectively activate PARP for participation in different cellular responses associated with different DNA lesions.

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“…In a recent paper Wang et al (2004) demonstrated in human cells that DDB2 is a downstream effector of p53, which recognizes CPDs and can stimulate XPC recruitment and binding at the site of damage (Fitch et al, 2003a). This provides a clear explanation for the defect in global repair in p53-deficient human cells: lack of ChIP assay for E2F1-3.…”

Section: Ddb2 Is Not Activated By P53mentioning

confidence: 99%

E2F regulates DDB2: consequences for DNA repair in Rb-deficient cells

Prost

Caldwell

et al. 2006

Oncogene

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DDB2, a gene mutated in XPE patients, is involved in global genomic repair especially the repair of cyclobutane pyrimidine dimers (CPDs), and is regulated by p53 in human cells. We show that DDB2 is expressed in mouse tissues and demonstrate, using primary mouse epithelial cells, that mouse DDB2 is regulated by E2F transcription factors. Retinoblastoma (Rb), a tumor suppressor critical for the control of cell cycle progression, regulates E2F activity. Using Cre-Lox technology to delete Rb in primary mouse hepatocytes, we show that DDB2 gene expression increases, leading to elevated DDB2 protein levels. Furthermore, we show that endogenous E2F1 and E2F3 bind to DDB2 promoter and that treatment with E2F1-antisense or E2F1-small interfering RNA (siRNA) decreases DDB2 transcription, demonstrating that E2F1 is a transcriptional regulator for DDB2. This has consequences for global genomic repair: in Rb-null cells, where E2F activity is elevated, global DNA repair is increased and removal of CPDs is more efficient than in wild-type cells. Treatment with DDB2-siRNA decreases DDB2 expression and abolishes the repair phenotype of Rb-null cells. In summary, these results identify a new regulatory pathway for DDB2 by E2F, which does not require but is potentiated by p53, and demonstrate that DDB2 is involved in global repair in mouse epithelial cells.

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p53 responsive nucleotide excision repair gene products p48 and XPC, but not p53, localize to sites of UV-irradiation-induced DNA damage, in vivo

Cited by 73 publications

References 49 publications

DNA Damage Binding Protein Component DDB1 Participates in Nucleotide Excision Repair through DDB2 DNA-binding and Cullin 4A Ubiquitin Ligase Activity

DNA Damage Binding Protein Component DDB1 Participates in Nucleotide Excision Repair through DDB2 DNA-binding and Cullin 4A Ubiquitin Ligase Activity

Mechanism of early biphasic activation of poly(ADP-ribose) polymerase-1 in response to ultraviolet B radiation

E2F regulates DDB2: consequences for DNA repair in Rb-deficient cells

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