The Involvement of Annexin II in Resistance to UVB-Induced Cell Death and in the Increased Nucleotide Excision Repair Capacity of UV-Damaged DNA in Human Cells

Nucleotide excision repair is the sole mechanism for removing the major UV photoproducts from genomic DNA in human cells. In vitro with human cell-free extract or purified excision repair factors, the damage is removed from naked DNA or nucleosomes in the form of 24- to 32-nucleotide-long oligomers (nominal 30-mer) by dual incisions. Whether the DNA damage is removed from chromatin in vivo in a similar manner and what the fate of the excised oligomer was has not been known previously. Here, we demonstrate that dual incisions occur in vivo identical to the in vitro reaction. Further, we show that transcription-coupled repair, which operates in the absence of the XPC protein, also generates the nominal 30-mer in UV-irradiated XP-C mutant cells. Finally, we report that the excised 30-mer is released from the chromatin in complex with the repair factors TFIIH and XPG. Taken together, our results show the congruence of in vivo and in vitro data on nucleotide excision repair in humans.

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Reversible Protein Phosphorylation Modulates Nucleotide Excision Repair of Damaged DNA by Human Cell Extracts

Nucleic Acids Research ◽

10.1093/nar/24.3.433 ◽

1996 ◽

Vol 24 (3) ◽

pp. 433-440 ◽

Cited By ~ 54

Author(s):

R. R. Ariza ◽

S. M. Keyse ◽

J. G. Moggs ◽

R. D. Wood

Keyword(s):

Protein Phosphorylation ◽

Nucleotide Excision Repair ◽

Human Cell ◽

Excision Repair ◽

Cell Extracts ◽

Nucleotide Excision ◽

Reversible Protein Phosphorylation ◽

Damaged Dna

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O V A.1 Nucleotide excision repair mechanism in human cells: Analysis by in vitro assays

Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis ◽

10.1016/s0027-5107(97)82703-0 ◽

1997 ◽

Vol 379 (1) ◽

pp. S33

Author(s):

Bernard Salles ◽

Patrick Calsou ◽

Philippe Frit ◽

Ruo-Ya Li ◽

Catherine Muller ◽

...

Keyword(s):

Nucleotide Excision Repair ◽

Excision Repair ◽

Human Cells ◽

In Vitro Assays ◽

Repair Mechanism ◽

Nucleotide Excision

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DNA damage and nucleotide excision repair capacity in healthy individuals

Environmental and Molecular Mutagenesis ◽

10.1002/em.20650 ◽

2011 ◽

Vol 52 (7) ◽

pp. 511-517 ◽

Cited By ~ 32

Author(s):

Jana Slyskova ◽

Alessio Naccarati ◽

Veronika Polakova ◽

Barbara Pardini ◽

Ludmila Vodickova ◽

...

Keyword(s):

Dna Damage ◽

Nucleotide Excision Repair ◽

Excision Repair ◽

Healthy Individuals ◽

Repair Capacity ◽

Nucleotide Excision

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Recruitment of the Nucleotide Excision Repair Endonuclease XPG to Sites of UV-Induced DNA Damage Depends on Functional TFIIH

Molecular and Cellular Biology ◽

10.1128/mcb.00695-06 ◽

2006 ◽

Vol 26 (23) ◽

pp. 8868-8879 ◽

Cited By ~ 56

Author(s):

Angelika Zotter ◽

Martijn S. Luijsterburg ◽

Daniël O. Warmerdam ◽

Shehu Ibrahim ◽

Alex Nigg ◽

...

Keyword(s):

Dna Damage ◽

Nucleotide Excision Repair ◽

Fluorescent Protein ◽

Core Protein ◽

Excision Repair ◽

Mammalian Cell Lines ◽

Nucleotide Excision ◽

Green Fluorescent ◽

Damaged Dna

ABSTRACT The structure-specific endonuclease XPG is an indispensable core protein of the nucleotide excision repair (NER) machinery. XPG cleaves the DNA strand at the 3′ side of the DNA damage. XPG binding stabilizes the NER preincision complex and is essential for the 5′ incision by the ERCC1/XPF endonuclease. We have studied the dynamic role of XPG in its different cellular functions in living cells. We have created mammalian cell lines that lack functional endogenous XPG and stably express enhanced green fluorescent protein (eGFP)-tagged XPG. Life cell imaging shows that in undamaged cells XPG-eGFP is uniformly distributed throughout the cell nucleus, diffuses freely, and is not stably associated with other nuclear proteins. XPG is recruited to UV-damaged DNA with a half-life of 200 s and is bound for 4 min in NER complexes. Recruitment requires functional TFIIH, although some TFIIH mutants allow slow XPG recruitment. Remarkably, binding of XPG to damaged DNA does not require the DDB2 protein, which is thought to enhance damage recognition by NER factor XPC. Together, our data present a comprehensive view of the in vivo behavior of a protein that is involved in a complex chromatin-associated process.

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Nucleotide Excision Repair or Polymerase V-Mediated Lesion Bypass Can Act To Restore UV-Arrested Replication Forks in Escherichia coli

Journal of Bacteriology ◽

10.1128/jb.187.20.6953-6961.2005 ◽

2005 ◽

Vol 187 (20) ◽

pp. 6953-6961 ◽

Cited By ~ 35

Author(s):

Charmain T. Courcelle ◽

Jerilyn J. Belle ◽

Justin Courcelle

Keyword(s):

Dna Synthesis ◽

Nucleotide Excision Repair ◽

Excision Repair ◽

Translesion Synthesis ◽

Replication Forks ◽

Repair Capacity ◽

Translesion Dna Synthesis ◽

Pol V ◽

Nucleotide Excision ◽

Translesion Dna

ABSTRACT Nucleotide excision repair and translesion DNA synthesis are two processes that operate at arrested replication forks to reduce the frequency of recombination and promote cell survival following UV-induced DNA damage. While nucleotide excision repair is generally considered to be error free, translesion synthesis can result in mutations, making it important to identify the order and conditions that determine when each process is recruited to the arrested fork. We show here that at early times following UV irradiation, the recovery of DNA synthesis occurs through nucleotide excision repair of the lesion. In the absence of repair or when the repair capacity of the cell has been exceeded, translesion synthesis by polymerase V (Pol V) allows DNA synthesis to resume and is required to protect the arrested replication fork from degradation. Pol II and Pol IV do not contribute detectably to survival, mutagenesis, or restoration of DNA synthesis, suggesting that, in vivo, these polymerases are not functionally redundant with Pol V at UV-induced lesions. We discuss a model in which cells first use DNA repair to process replication-arresting UV lesions before resorting to mutagenic pathways such as translesion DNA synthesis to bypass these impediments to replication progression.

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