Structural Insights into the Recognition of N 2 -Aryl- and C8-Aryl DNA Lesions by the Repair Protein XPA/Rad14

ChemBioChem ◽  
2017 ◽  
Vol 18 (14) ◽  
pp. 1379-1382 ◽  
Author(s):  
Charlotte Ebert ◽  
Nina Simon ◽  
Sabine Schneider ◽  
Thomas Carell
Keyword(s):  
Dna Lesions ◽  
Repair Protein ◽  
Structural Insights

scholarly journals The retinoblastoma homolog RBR 1 mediates localization of the repair protein RAD 51 to DNA lesions in Arabidopsis

2017 ◽  
Vol 36 (9) ◽  
pp. 1279-1297 ◽  
Author(s):  
Sascha Biedermann ◽  
Hirofumi Harashima ◽  
Poyu Chen ◽  
Maren Heese ◽  
Daniel Bouyer ◽  
...  
Keyword(s):  
Dna Lesions ◽  
Repair Protein

scholarly journals Repair protein persistence at DNA lesions characterizes XPF defect with Cockayne syndrome features

2018 ◽  
Vol 46 (18) ◽  
pp. 9563-9577 ◽  
Author(s):  
Mariangela Sabatella ◽  
Arjan F Theil ◽  
Cristina Ribeiro-Silva ◽  
Jana Slyskova ◽  
Karen Thijssen ◽  
...  
Keyword(s):  
Cockayne Syndrome ◽  
Dna Lesions ◽  
Repair Protein

Distinct kinetics of DNA repair protein accumulation at DNA lesions and cell cycle-dependent formation of γH2AX- and NBS1-positive repair foci

2015 ◽  
Vol 107 (12) ◽  
pp. 440-454 ◽  
Author(s):  
Jana Suchánková ◽  
Stanislav Kozubek ◽  
Soňa Legartová ◽  
Petra Sehnalová ◽  
Thomas Küntziger ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Dna Repair ◽  
Dna Lesions ◽  
Protein Accumulation ◽  
Repair Protein ◽  
Dna Repair Protein ◽  
Cycle Dependent ◽  
Kinetics Of

scholarly journals Two human homologs of Rad23 are functionally interchangeable in complex formation and stimulation of XPC repair activity.

1997 ◽  
Vol 17 (12) ◽  
pp. 6924-6931 ◽  
Author(s):  
K Sugasawa ◽  
J M Ng ◽  
C Masutani ◽  
T Maekawa ◽  
A Uchida ◽  
...  
Keyword(s):  
Excision Repair ◽  
Early Stage ◽  
Dna Lesions ◽  
Repair Protein ◽  
Nucleotide Excision ◽  
Repair Activity ◽  
In Vitro Repair ◽  
Stimulation Of ◽  
Active Genes

XPC-hHR23B protein complex is specifically involved in nucleotide excision repair (NER) of DNA lesions on transcriptionally inactive sequences as well as the nontranscribed strand of active genes. Here we demonstrate that not only highly purified recombinant hHR23B (rhHR23B) but also a second human homolog of the Saccharomyces cerevisiae Rad23 repair protein, hHR23A, stimulates the in vitro repair activity of recombinant human XPC (rhXPC), revealing functional redundancy between these human Rad23 homologs. Coprecipitation experiments with His-tagged rhHR23 as well as sedimentation velocity analysis showed that both rhHR23 proteins in vitro reconstitute a physical complex with rhXPC. Both complexes were more active than free rhXPC, indicating that complex assembly is required for the stimulation. rhHR23B was shown to stimulate an early stage of NER at or prior to incision. Furthermore, both rhHR23 proteins function in a defined NER system reconstituted with purified proteins, indicating direct involvement of hHR23 proteins in the DNA repair reaction via interaction with XPC.

scholarly journals Bypassing a 8,5′-cyclo-2′-deoxyadenosine lesion by human DNA polymerase η at atomic resolution

2018 ◽  
Vol 115 (42) ◽  
pp. 10660-10665 ◽  
Author(s):  
Peter J. Weng ◽  
Yang Gao ◽  
Mark T. Gregory ◽  
Pengcheng Wang ◽  
Yinsheng Wang ◽  
...  
Keyword(s):  
Excision Repair ◽  
Dna Lesions ◽  
Dna Structures ◽  
Extra Space ◽  
Human Dna ◽  
Nucleotide Excision ◽  
Mutagenic Property ◽  
Structural Insights ◽  
Protein Environment ◽  
Adenine Base

Oxidatively induced DNA lesions 8,5′-cyclopurine-2′-deoxynucleosides (cdPus) are prevalent and cytotoxic by impeding DNA replication and transcription. Both the 5′R- and 5′S-diastereomers of cdPu can be removed by nucleotide excision repair; however, the 5′S-cdPu is more resistant to repair than the 5′R counterpart. Here, we report the crystal structures of human polymerase (Pol) η bypassing 5′S-8,5′-cyclo-2′-deoxyadenosine (cdA) in insertion and the following two extension steps. The cdA-containing DNA structures vary in response to the protein environment. Supported by the “molecular splint” of Pol η, the structure of 5′S-cdA at 1.75-Å resolution reveals that the backbone is pinched toward the minor groove and the adenine base is tilted. In the templating position, the cdA takes up the extra space usually reserved for the thymine dimer, and dTTP is efficiently incorporated by Pol η in the presence of Mn2+. Rigid distortions of the DNA duplex by cdA, however, prevent normal base pairing and hinder immediate primer extension by Pol η. Our results provide structural insights into the strong replication blockage effect and the mutagenic property of the cdPu lesions in cells.

scholarly journals Quantitating repair protein accumulation at DNA lesions: Past, present, and future

DNA Repair ◽  
2019 ◽  
Vol 81 ◽  
pp. 102650 ◽  
Author(s):  
Jyothi Mahadevan ◽  
Samuel Bowerman ◽  
Karolin Luger
Keyword(s):  
Dna Lesions ◽  
Protein Accumulation ◽  
Repair Protein

scholarly journals Structural insights into the recognition of cisplatin and AAF-dG lesion by Rad14 (XPA)

2015 ◽  
Vol 112 (27) ◽  
pp. 8272-8277 ◽  
Author(s):  
Sandra C. Koch ◽  
Jochen Kuper ◽  
Karola L. Gasteiger ◽  
Nina Simon ◽  
Ralf Strasser ◽  
...  
Keyword(s):  
Excision Repair ◽  
Dna Lesions ◽  
General Mechanism ◽  
Dna Duplexes ◽  
Central Importance ◽  
Dna Structures ◽  
Cancer Predisposition Syndrome ◽  
Recognition Motif ◽  
Nucleotide Excision ◽  
Structural Insights

Nucleotide excision repair (NER) is responsible for the removal of a large variety of structurally diverse DNA lesions. Mutations of the involved proteins cause the xeroderma pigmentosum (XP) cancer predisposition syndrome. Although the general mechanism of the NER process is well studied, the function of the XPA protein, which is of central importance for successful NER, has remained enigmatic. It is known, that XPA binds kinked DNA structures and that it interacts also with DNA duplexes containing certain lesions, but the mechanism of interactions is unknown. Here we present two crystal structures of the DNA binding domain (DBD) of the yeast XPA homolog Rad14 bound to DNA with either a cisplatin lesion (1,2-GG) or an acetylaminofluorene adduct (AAF-dG). In the structures, we see that two Rad14 molecules bind to the duplex, which induces DNA melting of the duplex remote from the lesion. Each monomer interrogates the duplex with a β-hairpin, which creates a 13mer duplex recognition motif additionally characterized by a sharp 70° DNA kink at the position of the lesion. Although the 1,2-GG lesion stabilizes the kink due to the covalent fixation of the crosslinked dG bases at a 90° angle, the AAF-dG fully intercalates into the duplex to stabilize the kinked structure.

scholarly journals Human MMS21/NSE2 Is a SUMO Ligase Required for DNA Repair

2005 ◽  
Vol 25 (16) ◽  
pp. 7021-7032 ◽  
Author(s):  
Patrick Ryan Potts ◽  
Hongtao Yu
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Ectopic Expression ◽  
Well Being ◽  
Dna Lesions ◽  
Wild Type ◽  
Sumo Ligase ◽  
Repair Protein ◽  
Dna Repair Protein ◽  
Induced Apoptosis

ABSTRACT DNA repair is required for the genomic stability and well-being of an organism. In yeasts, a multisubunit complex consisting of SMC5, SMC6, MMS21/NSE2, and other non-SMC proteins is required for DNA repair through homologous recombination. The yeast MMS21 protein is a SUMO ligase. Here we show that the human homolog of MMS21 is also a SUMO ligase. hMMS21 stimulates sumoylation of hSMC6 and the DNA repair protein TRAX. Depletion of hMMS21 by RNA interference (RNAi) sensitizes HeLa cells toward DNA damage-induced apoptosis. Ectopic expression of wild-type hMMS21, but not its ligase-inactive mutant, rescues this hypersensitivity of hMMS21-RNAi cells. ATM/ATR are hyperactivated in hMMS21-RNAi cells upon DNA damage. Consistently, hMMS21-RNAi cells show an increased number of phospho-CHK2 foci. Finally, we show that hMMS21-RNAi cells show a decreased capacity to repair DNA lesions as measured by the comet assay. Our findings suggest that the human SMC5/6 complex and the SUMO ligase activity of hMMS21 are required for the prevention of DNA damage-induced apoptosis by facilitating DNA repair in human cells.

SAC3B is a target of CML19, the centrin 2 of Arabidopsis thaliana

2020 ◽  
Vol 477 (1) ◽  
pp. 173-189 ◽  
Author(s):  
Marco Pedretti ◽  
Carolina Conter ◽  
Paola Dominici ◽  
Alessandra Astegno
Keyword(s):  
Excision Repair ◽  
Complex Structure ◽  
Binding Motif ◽  
C Terminus ◽  
Repair Protein ◽  
Nucleotide Excision ◽  
Ef Hand ◽  
Molecular Determinants ◽  
Structure In Solution

Arabidopsis centrin 2, also known as calmodulin-like protein 19 (CML19), is a member of the EF-hand superfamily of calcium (Ca2+)-binding proteins. In addition to the notion that CML19 interacts with the nucleotide excision repair protein RAD4, CML19 was suggested to be a component of the transcription export complex 2 (TREX-2) by interacting with SAC3B. However, the molecular determinants of this interaction have remained largely unknown. Herein, we identified a CML19-binding site within the C-terminus of SAC3B and characterized the binding properties of the corresponding 26-residue peptide (SAC3Bp), which exhibits the hydrophobic triad centrin-binding motif in a reversed orientation (I8W4W1). Using a combination of spectroscopic and calorimetric experiments, we shed light on the SAC3Bp–CML19 complex structure in solution. We demonstrated that the peptide interacts not only with Ca2+-saturated CML19, but also with apo-CML19 to form a protein–peptide complex with a 1 : 1 stoichiometry. Both interactions involve hydrophobic and electrostatic contributions and include the burial of Trp residues of SAC3Bp. However, the peptide likely assumes different conformations upon binding to apo-CML19 or Ca2+-CML19. Importantly, the peptide dramatically increases the affinity for Ca2+ of CML19, especially of the C-lobe, suggesting that in vivo the protein would be Ca2+-saturated and bound to SAC3B even at resting Ca2+-levels. Our results, providing direct evidence that Arabidopsis SAC3B is a CML19 target and proposing that CML19 can bind to SAC3B through its C-lobe independent of a Ca2+ stimulus, support a functional role for these proteins in TREX-2 complex and mRNA export.

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