Heat-labile uracil-DNA glycosylase: purification and characterization.

FEBS Lett

Boehringer Mannheim GmbH, Penzberg, Germany.

Published: June 1996

A uracil-DNA glycosylase (UNG) from a psychrophilic marine bacterium (BMTU 3346) has been purified to apparent homogeneity. The enzyme has a molecular weight of 23400 Da. It is stable in complex buffers (containing glycerol/BSA), whereas it is heat-labile in dilute buffers (free of stabilizers) with a half-life of 2 min at 40 degrees C. Due to the thermolability, uracil-DNA glycosylase is suitable for application in the carryover prevention technique showing less residual activity and/or a slower reactivation rate than the usually applied UNG from Escherichia coli.

Download full-text PDF

Source
http://dx.doi.org/10.1016/0014-5793(96)00444-9DOI Listing

Publication Analysis

Top Keywords

uracil-dna glycosylase
12
heat-labile uracil-dna
4
glycosylase purification
4
purification characterization
4
characterization uracil-dna
4
glycosylase ung
4
ung psychrophilic
4
psychrophilic marine
4
marine bacterium
4
bacterium bmtu
4

Similar Publications

The kinetics of uracil-N-glycosylase distribution inside replication foci.

Sci Rep

January 2025

Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacký University Olomouc, Olomouc, Czech Republic.

Mismatched nucleobase uracil is commonly repaired through the base excision repair initiated by DNA uracil glycosylases. The data presented in this study strongly indicate that the nuclear uracil-N-glycosylase activity and nuclear protein content in human cell lines is highest in the S phase of the cell cycle and that its distribution kinetics partially reflect the DNA replication activity in replication foci. In this respect, the data demonstrate structural changes of the replication focus related to the uracil-N-glycosylase distribution several dozens of minutes before end of its replication.

View Article and Find Full Text PDF

Base deamination can lead to DNA base damage, among which cytosine deamination to uracil occurs frequently. Before repair, replication of uracil in DNA will generate GC → AT transversion mutation. Since base deamination is accelerated by high temperature, genomic DNA stability of hyperthermophiles, which grow optimally above 75 °C, is facing a severe threat by the elevated base deamination created by their living high temperature environments.

View Article and Find Full Text PDF

Targeted deaminase-free T-to-G and C-to-K base editing in rice by fused human uracil DNA glycosylase variants.

Plant Biotechnol J

January 2025

Institute of Crop Sciences/National Nanfan Research Institute, Chinese Academy of Agricultural Sciences (CAAS), and Key Laboratory of Gene Editing Technologies (Hainan), Ministry of Agriculture and Rural Affairs, Sanya, China.

View Article and Find Full Text PDF

The cytotoxic mechanisms of thymidylate synthase inhibitors, such as the multitarget antifolate pemetrexed, are not yet fully understood. Emerging evidence indicates that combining pemetrexed with histone deacetylase inhibitors (HDACi) may enhance therapeutic efficacy in non-small cell lung cancer (NSCLC). To explore this further, A549 NSCLC cells were treated with various combinations of pemetrexed and the HDACi MS275 (Entinostat), and subsequently assessed for cell viability, cell cycle changes, and genotoxic markers.

View Article and Find Full Text PDF

Genome-Wide A → G and C → T Mutations Induced by Functional TadA Variants in .

ACS Synth Biol

January 2025

Laboratory of Synthetic Microbiology, School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, P. R. China.

The fusion expression of deoxyribonucleic acid (DNA) replication-related proteins with nucleotide deaminase enzymes promotes random mutations in bacterial genomes, thereby increasing genetic diversity among the population. Most previous studies have focused on cytosine deaminase, which produces only C → T mutations, significantly limiting the variety of mutation types. In this study, we developed a fusion expression system by combining DnaG (RNA primase) with adenine deaminase TadA-8e (DnaG-TadA) in , which is capable of rapidly introducing A → G mutations into the genome, resulting in a 664-fold increase in terms of mutation rate.

View Article and Find Full Text PDF

Want 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!