A5014297467- DNA Repair Mechanisms
- Cardiomyopathy and Myosin Studies
- DNA and Nucleic Acid Chemistry
- Carcinogens and Genotoxicity Assessment
- Plant Genetic and Mutation Studies
- Muscle Physiology and Disorders
- Cellular Mechanics and Interactions
- Botulinum Toxin and Related Neurological Disorders
- CRISPR and Genetic Engineering
- RNA Research and Splicing
- Bacterial Genetics and Biotechnology
- Protein Structure and Dynamics
- Acute Ischemic Stroke Management
- Genomics and Chromatin Dynamics
- Bacteriophages and microbial interactions
- Cardiovascular Effects of Exercise
- Stroke Rehabilitation and Recovery
- RNA Interference and Gene Delivery
- Parkinson's Disease Mechanisms and Treatments
- Viral Infectious Diseases and Gene Expression in Insects
- Ion channel regulation and function
- Cerebral Palsy and Movement Disorders
- Neurogenetic and Muscular Disorders Research
- Dialysis and Renal Disease Management
- Genetics, Aging, and Longevity in Model Organisms
Eötvös Loránd University
2012-2022
Institute of Molecular Life Sciences
2013-2022
Motorpharma (Hungary)
2020-2022
HUN-REN Research Centre for Natural Sciences
2021
Printnet (Hungary)
2021
Western New England University
2021
Plant Protection Institute
2021
Centre for Agricultural Research
2021
Hungarian Academy of Sciences
2012-2017
University of California, Davis
2014
Formation of RAD51 filaments on single-stranded DNA is an essential event during homologous recombination, which required for homology search, strand exchange and protection replication forks. nucleoprotein (NF) development genomic stability, its failure associated with developmental abnormalities tumorigenesis. Here we describe the structure human NFs Walker box mutants using electron microscopy. Wild-type adopt 'open' conformation when compared to a 'closed' formed by mutants, reflecting...
Cells must continuously repair inevitable DNA damage while avoiding the deleterious consequences of imprecise repair. Distinction between legitimate and illegitimate processes is thought to be achieved in part through differential recognition processing specific noncanonical structures, although mechanistic basis discrimination remains poorly defined. Here, we show that Escherichia coli RecQ, a central recombination enzyme, exhibits substrates based on their geometry structure. Through...
The rate-limiting step of the myosin basal ATPase (i.e. in absence actin) is assumed to be a post-hydrolysis swinging lever arm (reverse recovery step), that limits subsequent rapid product release steps. However, direct experimental evidence for this assignment lacking. To investigate binding and ADP phosphate independently from motion, two single tryptophan-containing motor domains Dictyostelium II were used. tryptophans W129+ W501+ constructs are located at entrance nucleotide pocket near...
The powerstroke of the myosin motor is basis cell division and bodily movement, but has eluded empirical description due to short lifetime low abundance intermediates during force generation. To gain insight into this process, we used well-established single-tryptophan pyrene fluorescent sensors electron microscopy characterize structural kinetic properties complexed with ADP blebbistatin, a widely inhibitor. We found that blebbistatin does not weaken tight actin binding myosin.ADP,...
BLM, one of the human RecQ helicases, plays a fundamental role in homologous recombination-based error-free DNA repair pathways, which require its translocation and unwinding activities. Although is essential vivo during processes it provides framework for more complex activities including strand separation nucleoprotein displacement, mechanism has not been resolved any helicase. Here, we present quantitative model monomeric form BLM along ssDNA. We show that performs at low adenosine...
Abstract Homologous recombination (HR) is a ubiquitous and efficient process that serves the repair of severe forms DNA damage generation genetic diversity during meiosis. HR can proceed via multiple pathways with different outcomes may aid or impair genome stability faithful inheritance, underscoring importance quality control. Human Bloom’s syndrome (BLM, RecQ family) helicase plays central roles in pathway selection control unexplored molecular mechanisms. Here we show BLM’s multi-domain...
The single-stranded DNA binding protein (SSB) of Escherichia coli plays essential roles in maintaining genome integrity by sequestering ssDNA and mediating processing pathways through interactions with DNA-processing enzymes. Despite its DNA-sequestering properties, SSB stimulates the activities some partners. One example is maintenance RecQ helicase. Here, we determine mechanistic details RecQ–SSB interaction using single-molecule magnetic tweezers rapid kinetic experiments. Our results...
Bloom's syndrome DNA helicase (BLM), a member of the RecQ family, is key player in homologous recombination (HR)-based error-free repair processes.During HR, BLM exerts various biochemical activities including single-stranded (ss) translocation, separation and annealing complementary strands, disruption complex structures (e.g.displacement loops) contributes to quality control HR via clearance Rad51 nucleoprotein filaments.We performed quantitative mechanistic analysis truncated constructs...
Maintenance of genome integrity is the major biological role RecQ-family helicases via their participation in homologous recombination (HR)-mediated DNA repair processes. RecQ exert functions by using free energy ATP hydrolysis for mechanical movement along tracks (translocation). In addition to importance translocation per se processes, knowledge its mechanism necessary understanding more complex translocation-based activities, including nucleoprotein displacement, strand separation...
Acute ischemic stroke treatment faces an unresolved obstacle as capillary reperfusion remains insufficient after thrombolysis and thrombectomy causing neuronal damage poor prognosis.Hypoxia-induced constriction is mediated by actomyosin contraction in precapillary smooth muscle cells (SMCs) therefore myosin-2 could be ideal target with potentially high impact on of capillaries.Methods: The inhibitor para-aminoblebbistatin (AmBleb) was tested isolated human rat arterioles to assess the effect...
Homologous recombination (HR) is a key process in the repair of double-stranded DNA breaks (DSBs) that can initiate cancer or cell death. Human Bloom's syndrome RecQ-family helicase (BLM) exerts complex activities to promote DSB while avoiding illegitimate HR. The oligomeric assembly state BLM has been unresolved aspect its activities. In this study we assessed structure and BLM, absence presence HR-intermediate structures, by using single-molecule visualization (electron microscopic atomic...
Active site loops that are conserved across superfamilies of myosins, kinesins, and G proteins play key roles in allosteric coupling NTP hydrolysis to interaction with track filaments or effector proteins. In this study, we investigated how the class-specific natural variation switch-2 active loop contributes motor function intracellular transporter myosin-5. We used single-molecule, rapid kinetic spectroscopic experiments semiempirical quantum chemical simulations show structure including a...
Quantitative determination of enzymatic rates, processivity and mechanochemical coupling is a key aspect in characterizing nucleotide triphosphate (NTP)-driven nucleic acid motor enzymes, for both basic research technological applications. Here, we present streamlined analytical method suitable the all functional parameters based on measurement NTP hydrolysis during interaction enzymes with track. The proposed utilizes features kinetic time courses that have not been addressed previous...
Blebbistatin, <i>para</i>-nitroblebbistatin (NBleb), and <i>para</i>-aminoblebbistatin (AmBleb) are highly useful tool compounds as they selectively inhibit the ATPase activity of myosin-2 family proteins. Despite medical importance drug targets, chemical optimization has not yet provided a promising lead for development because previous structure-activity-relationship studies were limited to single isoform. Here we evaluated potential blebbistatin scaffold found that D-ring substitutions...
Myosin interacts with actin during its enzymatic cycle, and stimulates myosin's ATPase activity. There are extensive interaction surfaces on both myosin. Several surface loops of myosin play different roles in actomyosin interaction. However, the functional role loop 4 binding is still ambiguous. We explored by either mutating conserved acidic group, Glu-365, to Gln (E365Q), or replacing entire three glycines (ΔAL) a Dictyostelium discoideum II motor domain (MD) containing single tryptophan...
Dipolar electron paramagnetic resonance (EPR) has been used to determine the conformation of myosin actin-binding cleft in various states actomyosin ATPase and binding actin. Comparison crystal structures isolated components microscopy actin–myosin complex suggested that between upper lower 50 kDa subdomains closes on The changes might be coupled state nucleotide active site provide communication ATP hydrolysis actin binding. In order detect these solution, we measured distances double...
Abstract Subcellular dynamics of non-muscle myosin 2 (NM2) is crucial for a broad-array cellular functions. To unveil mechanisms NM2 pharmacological control, we determined how the diffusion affected by NM2′s allosteric inhibitors, i.e. blebbistatin derivatives, as compared to Y-27632 inhibiting ROCK, upstream regulator. We found that markedly faster in central fibers than peripheral stress fibers. accelerated both and fibers, whereas derivatives slightly at low, but slowed it high inhibitor...
The processing of various DNA structures by RecQ helicases is crucial for genome maintenance in both bacteria and eukaryotes. perform active destabilization duplexes, based on tight coupling their ATPase activity to moderately processive translocation along strands. Here, we determined the kinetic mechanism E. coli helicase reveal how mechanoenzymatic achieved. We found that interaction with results a drastic acceleration rate-limiting ATP cleavage step, which occurs productively due...