A5047138595- DNA Repair Mechanisms
- CRISPR and Genetic Engineering
- DNA and Nucleic Acid Chemistry
- Carcinogens and Genotoxicity Assessment
- Ubiquitin and proteasome pathways
- Genomics and Chromatin Dynamics
- RNA modifications and cancer
- Cancer-related Molecular Pathways
- Skin Protection and Aging
- Monoclonal and Polyclonal Antibodies Research
- Cancer therapeutics and mechanisms
- Peptidase Inhibition and Analysis
- Genetics and Neurodevelopmental Disorders
- RNA Research and Splicing
- Cell death mechanisms and regulation
- Genomics, phytochemicals, and oxidative stress
- bioluminescence and chemiluminescence research
- Polyomavirus and related diseases
- Melanoma and MAPK Pathways
- Spaceflight effects on biology
- Cancer Research and Treatments
- Prenatal Screening and Diagnostics
- Click Chemistry and Applications
- interferon and immune responses
- Porphyrin Metabolism and Disorders
Kanazawa University
2007-2023
University of California, San Francisco
2008
University of North Carolina at Chapel Hill
1997-1999
Indiana University School of Medicine
1997-1999
Damaged DNA-binding protein, DDB, is a heterodimer of p127 and p48 with high specificity for binding to several types DNA damage. Mutations in the <i>p48</i> gene that cause loss DDB activity were found subset xeroderma pigmentosum complementation group E (XP-E) patients have linked deficiency global genomic repair cyclobutane pyrimidine dimers (CPDs) these cells. Here we show highly defined system purified factors, can greatly stimulate excision reaction reconstituted XPA, RPA, XPC·HR23B,...
Human excision nuclease removes DNA damage by concerted dual incisions bracketing the lesion. The are accomplished sequential and partly overlapping actions of six repair factors, RPA, XPA, XPC, TFIIH, XPG, XPF·ERCC1. Of these, XPC have specific binding affinity for damaged DNA. To learn about role these three proteins in recognition order assembly nuclease, we measured affinities to a fragment containing single (6-4) photoproduct determined rate under variety reaction conditions. We found...
The assembly and composition of human excision nuclease were investigated by electrophoretic mobility shift assay DNase I footprinting. Individual repair factors or any combination up to four failed form DNA–protein complexes high specificity stability. A stable complex can be detected only when XPA/RPA, transcription factor IIH, XPC⋅HHR23B, XPG ATP are present in the reaction mixture. XPF⋅ERCC1 heterodimer changes formed with other five factors, but it does not confer additional...
Nucleotide excision repair in humans is a complex reaction involving 14 polypeptides six factors for dual incisions on either sides of DNA lesion. To identify the intermediates that form by human nuclease, we adopted three approaches: purification functional DNA.protein complexes, permanganate footprinting, and employment as substrate presumptive containing unwound sequences 5' to, 3' or encompassing The first detectable intermediate was formed binding XPA, RPA, XPC.HHR23B plus TFIIH...
XPG is a member of the FEN-1 structure-specific endonuclease family. It has 3′-junction cutting activity on bubble substrates and makes 3′-incision in human dual incision (excision nuclease) repair system. To investigate precise role nucleotide excision repair, we mutagenized two amino acid residues thought to be involved DNA binding catalysis, overproduced mutant proteins using baculovirus/insect cell system, purified characterized proteins. The mutation D77A had modest effect junction gave...
Human cells contain a protein that binds to UV-irradiated DNA with high affinity. This protein, damaged DNA-binding (DDB), is heterodimer of two polypeptides, p127 and p48. Recent in vivo studies suggested DDB involved global genome repair cyclobutane pyrimidine dimers (CPDs), but the mechanism remains unclear. Here, we show vitro directly stimulates excision CPDs not (6-4)photoproducts. The activity cell-free extracts from Chinese hamster AA8 cell line lacks was increased 3-4-fold by...
Abstract Accumulating evidence indicates that transcription is closely related to DNA damage formation and the loss of RNA biogenesis factors causes genome instability. However, whether such are involved in responses remains unclear. We focus here on helicase Aquarius ( AQR ), a known R-loop processing factor, show its depletion human cells results accumulation during S phase, mediated by formation. investigated involvement found knockdown decreased damage-induced foci Rad51 replication...
Significance NER removes helix-destabilizing bulky adducts including UV lesions and crosslinks generated by formaldehyde. protects skin cells from carcinogenesis. XP patients deficient in develop progressive neurological dysfunction. We discovered that collaboration between TOP1 the BER pathway can substitute for lack of NER. This previously unappreciated repair is initiated TOP1-dependent sensing topological alterations near subsequent SSB formation followed BER. Considering rate-limiting...
The Chk1 kinase is highly conserved from yeast to humans and well known function in the cell cycle checkpoint induced by genotoxic or replication stress. activation of achieved ATR-dependent phosphorylation with aid additional factors. Robust insults induce apoptosis instead checkpoint, some components ATR-Chk1 pathway are cleaved active caspases, although it has been unclear whether attenuation role induction. Here we show that activated caspase-dependent cleavage when cells undergo...
Damaged DNA-binding protein (DDB), consisting of DDB1 and DDB2 subunits recognizes a wide spectrum DNA lesions. DDB is dispensable for in vitro nucleotide excision repair (NER) reaction, but stimulates this reaction especially cyclobutane pyrimidine dimer (CPD). Here we show that directly interacts with XPA, one core NER factors, mainly through subunit the amino-acid residues between 185 226 XPA are important interaction. Interestingly, point mutation causing substitution from Arg-207 to...
Replicative DNA polymerases are frequently stalled at damaged template strands. Stalled replication forks restored by the damage tolerance (DDT) pathways, error-prone translesion synthesis (TLS) to cope with excessive damage, and error-free switching (TS) homologous recombination. PDIP38 (Pol-delta interacting protein of 38 kDa), also called Pol δ-interacting 2 (PolDIP2), physically associates TLS polymerases, polymerase η (Polη), Polλ, PrimPol, activates them in vitro. It remains unclear...
To analyze the function of xeroderma pigmentosum group A (XPA) protein in strand-specific DNA repair, we examined repair UV-induced cyclobutane pyrimidine dimer (CPD) transcribed and non-transcribed strands dihydrofolate reductase gene (XP-A) cell line (XP12ROSV) which was transfected with various types mutant XPA cDNA. The transfectant overexpressing a defect interaction either ERCC1, replication (RPA), or general transcription factor TFIIH, showed more less decreased CPD each strand...
Abstract The multisubunit complex transcription factor IIH (TFIIH) has dual functions in transcriptional initiation and nucleotide excision repair (NER). TFIIH is comprised of two subcomplexes, the core subcomplex (seven subunits) including XPB XPD helicases cyclin‐dependent kinase (CDK)‐activating (CAK) (three containing CDK7 kinase. Recently, it been reported that spironolactone, an anti‐aldosterone drug, inhibits cellular NER by inducing proteasomal degradation potentiates cytotoxicity...
DNA photolesions induced by UV, cyclobutane pyrimidine dimer (CPD) and (6-4) photoproduct (6-4PP), are repaired nucleotide excision repair (NER) in human cells. Various immunoassays using monoclonal antibodies specific for the have been developed widely used analysis of cellular NER activity. In this study, we newly a microplate-formatted cell-based immunoassay, based on indirect immunofluorescence staining with lesion-specific combined an infrared imaging system. Using assay, show kinetics...
The ultraviolet B (UVB) component of sunlight can cause severe damage to skin cells and even induce cancer. Growing evidence indicates that the UVB-induced signaling network is complex involves diverse cellular processes. In this study, we investigated role c-Jun NH2 -terminal kinase-associated leucine zipper protein (JLP), a scaffold for mitogen-activated kinase (MAPK) cascades, in apoptosis. We found epidermal apoptosis was prevented Jlp knockout (KO) as well keratinocyte-specific KO mice....