Modifications of the comet assay were introduced to measure crosslinks and the effect of formaldehyde in liver cells of the tested animals was investigated by the new method to see whether the method is feasible. Since exposure of slides to proteinase K can increase DNA migration of the treated cells, the presence of DNA-protein crosslinks can be indicated by compare of the tail moment before and after the proteinase K added. The results showed that the modified protocol of the alkaline comet assay is a fast, inexpensive and sensitive tool for the detection of potent crosslinkers-induced DNA-protein crosslinks at single cell level. Due to its specific advantages, the modified comet assay seems to be a useful tool as a DNA crosslink potency indicator.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.3724/sp.j.1005.2008.00659 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Reframing Formalin: A Molecular Opportunity Enabling Historical Epigenomics and Retrospective Gene Expression Studies.
Mol Ecol Resour
January 2025
National Research Collections Australia, Commonwealth Scientific Industrial Research Organisation, Canberra, Australian Capital Territory, Australia.
Formalin preservation of museum specimens has long been considered a barrier to molecular research due to extensive crosslinking and chemical modification. However, recent optimisation of hot alkaline lysis and proteinase K digestion DNA extraction methods have enabled a growing number of studies to overcome these challenges and conduct genome-wide re-sequencing and targeted locus-specific sequencing. The newest, and perhaps most unexpected utility of formalin preservation in archival samples is its ability to preserve in situ DNA-protein interactions at a molecular level.
View Article and Find Full Text PDFCytidine analogs in plant epigenetic research and beyond.
J Exp Bot
December 2024
Centre of Plant Structural and Functional Genomics, Institute of Experimental Botany, Czech Acad Sci, Šlechtitelů 31, Olomouc 77900, Czech Republic.
Cytosine (DNA) methylation plays important roles in silencing transposable elements, plant development, genomic imprinting, stress responses, and maintenance of genome stability. To better understand the functions of this epigenetic modification, several tools have been developed to manipulate DNA methylation levels. These include mutants of DNA methylation writers and readers, targeted manipulation of locus-specific methylation, and the use of chemical inhibitors.
View Article and Find Full Text PDFSPRTN metalloprotease participates in repair of ROS-mediated DNA-protein crosslinks.
Sci Rep
December 2024
Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN, 55455, USA.
Exposure to reactive oxygen species (ROS) can induce DNA-protein crosslinks (DPCs), unusually bulky DNA lesions that block replication and transcription and play a role in aging, cancer, cardiovascular disease, and neurodegenerative disorders. Repair of DPCs depends on the coordinated efforts of proteases and DNA repair enzymes to cleave the protein component of the lesion to smaller DNA-peptide crosslinks which can be processed by tyrosyl-DNA phosphodiesterases 1 and 2, nucleotide excision and homologous recombination repair pathways. DNA-dependent metalloprotease SPRTN plays a role in DPC repair, and SPRTN-deficient mice exhibit an accelerated aging phenotype and develop liver cancer early in life.
View Article and Find Full Text PDFMultiple DNA repair pathways prevent acetaldehyde-induced mutagenesis in yeast.
Genetics
December 2024
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC 29425, USA.
Acetaldehyde is the primary metabolite of alcohol and is present in many environmental sources including tobacco smoke. Acetaldehyde is genotoxic, whereby it can form DNA adducts and lead to mutagenesis. Individuals with defects in acetaldehyde clearance pathways have increased susceptibility to alcohol-associated cancers.
View Article and Find Full Text PDFTranscription-coupled repair - mechanisms of action, regulation, and associated human disorders.
FEBS Lett
December 2024
Department of Genetics, Research Institute of Environmental Medicine (RIeM), Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8601, Japan.
The transcription-coupled repair (TCR) pathway resolves transcription-blocking DNA lesions to maintain cellular function and prevent transcriptional arrest. Stalled RNA polymerase II (RNAPII) triggers repair mechanisms, including RNAPII ubiquitination, which recruit UVSSA and TFIIH. Defects in TCR-associated genes cause disorders like Cockayne syndrome, UV-sensitive syndrome, xeroderma pigmentosum, and recently defined AMeDS.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!