scholarly journals Reconstitution of Yeast Nucleotide Excision Repair with Purified Rad Proteins, Replication Protein A, and Transcription Factor TFIIH

1995 ◽  
Vol 270 (22) ◽  
pp. 12973-12976 ◽  
Author(s):  
Sami N. Guzder ◽  
Yvette Habraken ◽  
Patrick Sung ◽  
Louise Prakash ◽  
Satya Prakash
Keyword(s):  
Transcription Factor ◽  
Nucleotide Excision Repair ◽  
Excision Repair ◽  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
Nucleotide Excision

scholarly journals DNA Replication but Not Nucleotide Excision Repair Is Required for UVC-Induced Replication Protein A Phosphorylation in Mammalian Cells

2000 ◽  
Vol 20 (8) ◽  
pp. 2696-2705 ◽  
Author(s):  
Gregory Rodrigo ◽  
Sophie Roumagnac ◽  
Marc S. Wold ◽  
Bernard Salles ◽  
Patrick Calsou
Keyword(s):  
Dna Damage ◽  
Dna Replication ◽  
Nucleotide Excision Repair ◽  
Mammalian Cells ◽  
Excision Repair ◽  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
Dependent Manner ◽  
Nucleotide Excision

ABSTRACT Exposure of mammalian cells to short-wavelength light (UVC) triggers a global response which can either counteract the deleterious effect of DNA damage by enabling DNA repair or lead to apoptosis. Several stress-activated protein kinases participate in this response, making phosphorylation a strong candidate for being involved in regulating the cellular damage response. One factor that is phosphorylated in a UVC-dependent manner is the 32-kDa subunit of the single-stranded DNA-binding replication protein A (RPA32). RPA is required for major cellular processes like DNA replication, and removal of DNA damage by nucleotide excision repair (NER). In this study we examined the signal which triggers RPA32 hyperphosphorylation following UVC irradiation in human cells. Hyperphosphorylation of RPA was observed in cells from patients with either NER or transcription-coupled repair (TCR) deficiency (A, C, and G complementation groups of xeroderma pigmentosum and A and B groups of Cockayne syndrome, respectively). This exclude both NER intermediates and TCR as essential signals for RPA hyperphosphorylation. However, we have observed that UV-sensitive cells deficient in NER and TCR require lower doses of UV irradiation to induce RPA32 hyperphosphorylation than normal cells, indicating that persistent unrepaired lesions contribute to RPA phosphorylation. Finally, the results of UVC irradiation experiments on nonreplicating cells and S-phase-synchronized cells emphasize a major role for DNA replication arrest in the presence of UVC lesions in RPA UVC-induced hyperphosphorylation in mammalian cells.

Differential binding kinetics of replication protein A during replication and the pre- and post-incision steps of nucleotide excision repair

DNA Repair ◽  
2014 ◽  
Vol 24 ◽  
pp. 46-56 ◽  
Author(s):  
Audrey M. Gourdin ◽  
Loes van Cuijk ◽  
Maria Tresini ◽  
Martijn S. Luijsterburg ◽  
Alex L. Nigg ◽  
...  
Keyword(s):  
Nucleotide Excision Repair ◽  
Excision Repair ◽  
Protein A ◽  
Replication Protein A ◽  
Binding Kinetics ◽  
Replication Protein ◽  
Nucleotide Excision ◽  
Differential Binding ◽  
Kinetics Of

scholarly journals Localization of xeroderma pigmentosum group A protein and replication protein A on damaged DNA in nucleotide excision repair

2010 ◽  
Vol 38 (22) ◽  
pp. 8083-8094 ◽  
Author(s):  
Yuliya S. Krasikova ◽  
Nadejda I. Rechkunova ◽  
Ekaterina A. Maltseva ◽  
Irina O. Petruseva ◽  
Olga I. Lavrik
Keyword(s):  
Nucleotide Excision Repair ◽  
Xeroderma Pigmentosum ◽  
Excision Repair ◽  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
Nucleotide Excision ◽  
Group A ◽  
Xeroderma Pigmentosum Group A ◽  
Damaged Dna

scholarly journals Persistently stalled replication forks inhibit nucleotide excision repair in trans by sequestering Replication protein A

2014 ◽  
Vol 42 (7) ◽  
pp. 4406-4413 ◽  
Author(s):  
Anastasia Tsaalbi-Shtylik ◽  
Jill Moser ◽  
Leon H. F. Mullenders ◽  
Jacob G. Jansen ◽  
Niels de Wind
Keyword(s):  
Nucleotide Excision Repair ◽  
Excision Repair ◽  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
Replication Forks ◽  
Nucleotide Excision ◽  
In Trans ◽  
Stalled Replication Forks

scholarly journals Relationship of the Xeroderma Pigmentosum Group E DNA Repair Defect to the Chromatin and DNA Binding Proteins UV-DDB and Replication Protein A

1998 ◽  
Vol 18 (6) ◽  
pp. 3182-3190 ◽  
Author(s):  
Vesna RapićOtrin ◽  
Isao Kuraoka ◽  
Tiziana Nardo ◽  
Mary McLenigan ◽  
A. P. M. Eker ◽  
...  
Keyword(s):  
Dna Binding ◽  
Nucleotide Excision Repair ◽  
Xeroderma Pigmentosum ◽  
Excision Repair ◽  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
Cell Extracts ◽  
Nucleotide Excision ◽  
Repair Defect

ABSTRACT Cells from complementation groups A through G of the heritable sun-sensitive disorder xeroderma pigmentosum (XP) show defects in nucleotide excision repair of damaged DNA. Proteins representing groups A, B, C, D, F, and G are subunits of the core recognition and incision machinery of repair. XP group E (XP-E) is the mildest form of the disorder, and cells generally show about 50% of the normal repair level. We investigated two protein factors previously implicated in the XP-E defect, UV-damaged DNA binding protein (UV-DDB) and replication protein A (RPA). Three newly identified XP-E cell lines (XP23PV, XP25PV, and a line formerly classified as an XP variant) were defective in UV-DDB binding activity but had levels of RPA in the normal range. The XP-E cell extracts did not display a significant nucleotide excision repair defect in vitro, with either UV-irradiated DNA or a uniquely placed cisplatin lesion used as a substrate. Purified UV-DDB protein did not stimulate repair of naked DNA by DDB− XP-E cell extracts, but microinjection of the protein into DDB−XP-E cells could partially correct the repair defect. RPA stimulated repair in normal, XP-E, or complemented extracts from other XP groups, and so the effect of RPA was not specific for XP-E cell extracts. These data strengthen the connection between XP-E and UV-DDB. Coupled with previous results, the findings suggest that UV-DDB has a role in the repair of DNA in chromatin.

scholarly journals An interaction between the DNA repair factor XPA and replication protein A appears essential for nucleotide excision repair.

1995 ◽  
Vol 15 (10) ◽  
pp. 5396-5402 ◽  
Author(s):  
L Li ◽  
X Lu ◽  
C A Peterson ◽  
R J Legerski
Keyword(s):  
Nucleotide Excision Repair ◽  
Excision Repair ◽  
Protein A ◽  
Simian Virus 40 ◽  
Replication Protein A ◽  
Simian Virus ◽  
Replication Protein ◽  
Nucleotide Excision

Replication protein A (RPA) is required for simian virus 40-directed DNA replication in vitro and for nucleotide excision repair (NER). Here we report that RPA and the human repair protein XPA specifically interact both in vitro and in vivo. Mapping of the RPA-interactive domains in XPA revealed that both of the largest subunits of RPA, RPA-70 and RPA-34, interact with XPA at distinct sites. A domain involved in mediating the interaction with RPA-70 was located between XPA residues 153 and 176. Deletion of highly conserved motifs within this region identified two mutants that were deficient in binding RPA in vitro and highly defective in NER both in vitro and in vivo. A second domain mediating the interaction with RPA-34 was identified within the first 58 residues in XPA. Deletion of this region, however, only moderately affects the complementing activity of XPA in vivo. Finally, the XPA-RPA complex is shown to have a greater affinity for damaged DNA than XPA alone. Taken together, these results indicate that the interaction between XPA and RPA is required for NER but that only the interaction with RPA-70 is essential.

scholarly journals DNA-binding polarity of human replication protein A positions nucleases in nucleotide excision repair

1998 ◽  
Vol 12 (16) ◽  
pp. 2598-2609 ◽  
Author(s):  
W. L. de Laat ◽  
E. Appeldoorn ◽  
K. Sugasawa ◽  
E. Weterings ◽  
N. G.J. Jaspers ◽  
...  
Keyword(s):  
Dna Binding ◽  
Nucleotide Excision Repair ◽  
Excision Repair ◽  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
Nucleotide Excision

scholarly journals Interactions of human replication protein A with single-stranded DNA adducts

2005 ◽  
Vol 385 (2) ◽  
pp. 519-526 ◽  
Author(s):  
Yiyong LIU ◽  
Zhengguan YANG ◽  
Christopher D. UTZAT ◽  
Yu LIU ◽  
Nicholas E. GEACINTOV ◽  
...  
Keyword(s):  
Excision Repair ◽  
Protein A ◽  
Chemical Properties ◽  
Replication Protein A ◽  
Replication Protein ◽  
Base Stacking ◽  
Single Stranded Dna ◽  
Protein Residues ◽  
Nucleotide Excision

Human RPA (replication protein A), a single-stranded DNA-binding protein, is required for many cellular pathways including DNA repair, recombination and replication. However, the role of RPA in nucleotide excision repair remains elusive. In the present study, we have systematically examined the binding of RPA to a battery of well-defined ssDNA (single-stranded DNA) substrates using fluorescence spectroscopy. These substrates contain adducts of (6-4) photoproducts, N-acetyl-2-aminofluorene-, 1-aminopyrene-, BPDE (benzo[a]pyrene diol epoxide)- and fluorescein that are different in many aspects such as molecular structure and size, DNA disruption mode (e.g. base stacking or non-stacking), as well as chemical properties. Our results showed that RPA has a lower binding affinity for damaged ssDNA than for non-damaged ssDNA and that the affinity of RPA for damaged ssDNA depends on the type of adduct. Interestingly, the bulkier lesions have a greater effect. With a fluorescent base-stacking bulky adduct, (+)-cis-anti-BPDE-dG, we demonstrated that, on binding of RPA, the fluorescence of BPDE-ssDNA was significantly enhanced by up to 8–9-fold. This indicated that the stacking between the BPDE adduct and its neighbouring ssDNA bases had been disrupted and there was a lack of substantial direct contacts between the protein residues and the lesion itself. For RPA interaction with short damaged ssDNA, we propose that, on RPA binding, the modified base of ssDNA is looped out from the surface of the protein, permitting proper contacts of RPA with the remaining unmodified bases.

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