Judit E Szabó

Hungarian Academy of Sciences | HAS · Institute of Enzymology

Fine-tuned regulation of the cellular nucleotide pools is indispensable for faithful replication of DNA. The genetic information is also safeguarded by DNA damage recognition and repair processes. Uracil is one of the most frequently occurring erroneous bases in DNA; it can arise from cytosine deamination or thymine-replacing incorporation. Two enz...

Transfer of phage-related pathogenicity islands of Staphylococcus aureus (SaPI-s) was recently reported to be activated by helper phage dUTPases. This is a novel function for dUTPases otherwise involved in preservation of genomic integrity by sanitizing the dNTP pool. Here we investigated the molecular mechanism of the dUTPase-induced gene expressi...

The regulation model of the Staphylococcus aureus pathogenicity island SaPIbov1 transfer was recently reported. The repressor protein Stl obstructs the expression of SaPI proteins Str and Xis, latter which is responsible for mobilization initiation. Upon Φ11 phage infection of S. aureus. phage dUTPase activates the SaPI transfer via Stl-dUTPase com...

Members of the dUTPase superfamily play an important role in the maintenance of the pyrimidine nucleotide balance and of genome integrity. dCTP deaminases and the bifunctional dCTP deaminase-dUTPases are cooperatively regulated by dTTP. However, the manifestation of allosteric behavior within the same trimeric protein architecture of dUTPases, the...

We investigated the potential (d)NDP/(d)NTP discrimination mechanisms in nucleotide pyrophosphatases. Here, we report that dUTPase, an essential nucleotide pyrophosphatase, uses a C-terminal P-loop-like sequence in a unique mechanism for substrate discrimination and efficient hydrolysis. Our spectroscopy and transient kinetics results on human dUTP...

Stimulated by the growing interest in the role of dNTP pools in physiological and malignant processes, we established dNTPpoolDB, the database that offers access to quantitative data on dNTP pools from a wide range of species, experimental and developmental conditions (https://dntppool.org/). The database includes measured absolute or relative cell...

Recently it was proposed that the redox status of cysteines acts as a redox switch to regulate both the oligomeric status and the activity of human dUTPase. In a separate report, a human dUTPase point mutation, resulting in a tyrosine to cysteine substitution (Y54C) was identified as the monogenic cause of a rare syndrome associated with diabetes a...

Cells maintain a fine-tuned, dynamic concentration balance in the pool of deoxyribonucleoside 5'-triphosphates (dNTPs). This balance is essential for physiological processes including cell cycle control or antiviral defense. Its perturbation results in increased mutation frequencies, replication arrest and may promote cancer development. An easily...

Clinical evidence shows that following initial response to treatment, drug-resistant cancer cells frequently evolve and, eventually, most tumors become resistant to all available therapies. We compiled a focused library consisting of >500 commercially available or newly synthetized 8-hydroxyquinoline (8OHQ) derivatives whose toxicity is paradoxical...

Protein inhibitors of key DNA repair enzymes play an important role in deciphering physiological pathways responsible for genome integrity, and may also be exploited in biomedical research. The staphylococcal repressor StlSaPIbov1 protein was described to be an efficient inhibitor of dUTPase homologues showing a certain degree of species-specificit...

Human deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase), essential for DNA integrity, acts as a survival factor for tumor cells and is a target for cancer chemotherapy. Here we report that the Staphylococcal repressor protein StlSaPIBov1(Stl) forms strong complex with human dUTPase. Functional analysis reveals that this interaction results...

List of prokaryotic genomes with the simultaneous lack of the dut and ung genes (dut–, ung– genotype). The table provides gives the list of the prokaryotic (bacterial/archaeal) genomes that lack both the dUTPase and UNG genes.

List of prokaryotic genomes where the dut gene is absent and the ung gene is present (dut–, ung+ genotype). The table provides gives the list of the prokaryotic (bacterial/archaeal) genomes without the dUTPase but with the UNG gene. The second column shows the presence of UNG inhibitors in the genome.

The distribution of bacterial/Archaeal genomes with and without dUTPase at the family level. Only those families are shown that have at least 15 genomes examined. Each node of the tree is labeled by three numbers: the first is the number of genomes with dUTPase under the node (lilac color on the pie graph segment); the second is the number of genom...

EMSA panels (A and B) of inter-L+tt and Stl site-tt, respectively. Stl concentration is shown in the top row; arrow indicates position of free DNA. (TIF)

EMSA panels (A, B and C) of Stl site-R poliA, Stl site-R G17T and Stl site-R C7G, respectively. Stl concentration is shown in the top row; arrow indicates position of free DNA. (TIF)

Schematic representation of the Stl binding sites and their relative locations of -10 and -35 boxes of the affected genes. The repressor or activator effects of the Stl protein exerted on to the respective genes are also indicated with blunt ended and arrow-headed lines, respectively. (TIF)

EMSA panels (A and B) of inter RR and inter RL, respectively. Stl concentration is shown in the top row; arrow indicates position of free DNA. (TIF)

EMSA panel of seq aspecific. Stl concentration is shown in the top row; arrow indicates position of free DNA. (TIF)

This document includes Supplementary Tables. Supplementary Table A lists the sequences of all oligonucleotides used in this study. Supplementary Table B presents the summary of the pairwase alignment of the stl-str and the str-xis intergenic regions, while Supplementary Table C provides a summary of the BPROM promoter prediction for stl, str and xi...

Fine-tuned regulation of the cellular nucleotide pools is indispensable for faithful replication of DNA. The genetic information is also safeguarded by DNA damage recognition and repair processes. Uracil is one of the most frequently occurring erroneous base in DNA; it can arise from cytosine deamination or thymine-replacing incorporation. Two enzy...

dUTPases are essential for maintaining genome integrity. Recently, in the case of a dUTPase from a Staphylococcal phage, another different physiological function was also suggested. Namely, it was shown that dUTPase from the Staphylococcus aureus bacteriophage 11 is capable of binding to the Staphylococcal Stl repressor protein. This binding interf...

Proteins responsible for the integrity of the genome are often used targets in drug therapies against various diseases. The inhibitors of these proteins are also important to study the pathways in genome integrity maintenance. A prominent example is Ugi, a well known cross-species inhibitor protein of the enzyme uracil-DNA glycosylase, responsible...

Genome integrity requires well controlled cellular pools of nucleotides. dUTPases are responsible for regulating cellular dUTP levels and providing dUMP for dTTP biosynthesis. In Staphylococcus , phage dUTPases are also suggested to be involved in a moonlighting function regulating the expression of pathogenicity-island genes. Staphylococcal phage...

Homologous recombination (HR) is essential for maintaining genomic integrity, which is challenged by a wide variety of potentially lethal DNA lesions. Regardless of the damage type, recombination is known to proceed by RAD51-mediated D-loop formation, followed by DNA repair synthesis. Nevertheless, the participating polymerases and extension mechan...

The phosphate-binding loop (P-loop) or Walker A sequence is a common feature of a large number of ATP and GTP binding proteins including kinases, cytoskeleton and DNA motors, membrane pumps and transporters. All known P-loop containing proteins are able to sense the difference between bound NTP and NDP via their P-loop which allows for hydrolyzing...