A5037348231- DNA Repair Mechanisms
- Cardiac electrophysiology and arrhythmias
- DNA and Nucleic Acid Chemistry
- Cancer therapeutics and mechanisms
- Genetic Neurodegenerative Diseases
- Mitochondrial Function and Pathology
- Ion channel regulation and function
- Cardiac Arrhythmias and Treatments
- Cardiac pacing and defibrillation studies
- Synthesis and bioactivity of alkaloids
- RNA and protein synthesis mechanisms
- Advanced biosensing and bioanalysis techniques
- Virus-based gene therapy research
- Electrochemical Analysis and Applications
- Atrial Fibrillation Management and Outcomes
- Fungal and yeast genetics research
- Biochemical and Molecular Research
- Neuroscience and Neural Engineering
- Congenital heart defects research
- RNA regulation and disease
- RNA Research and Splicing
- HIV Research and Treatment
- Cardiac Arrest and Resuscitation
- Corrosion Behavior and Inhibition
- Nanopore and Nanochannel Transport Studies
Centro de Investigación en Red en Enfermedades Cardiovasculares
2020-2024
Hospital General Universitario Gregorio Marañón
2020-2024
Universidad Complutense de Madrid
2020-2024
Centro de Investigación Biomédica en Red
2022-2024
Centro de Investigaciones Biológicas Margarita Salas
2014-2024
Centro de Investigación Biomédica en Red de Cáncer
2024
Instituto de Salud Carlos III
2023
Tufts University
2018-2023
Consejo Superior de Investigaciones Científicas
2015
Institut Cochin
1996
In a family with inappropriate sinus tachycardia (IST), we identified mutation (p.V240M) of the hyperpolarization-activated cyclic nucleotide–gated type 4 (HCN4) channel, which contributes to pacemaker current (I f ) in human sinoatrial node cells. Here, clinically study fifteen members and functionally analyze p.V240M variant. Macroscopic HCN4 single-channel currents were recorded using patch-clamp cells expressing native (WT) and/or channels. All carriers exhibited IST that was accompanied...
Friedreich's ataxia (FRDA) is caused by expansions of GAA•TTC repeats in the first intron human FXN gene that occur during both intergenerational transmissions and somatic cells. Here we describe an experimental system to analyze large-scale repeat cultured It employs a shuttle plasmid can replicate from SV40 origin cells or be stably maintained S. cerevisiae utilizing ARS4-CEN6. also contains selectable cassette allowing us detect accumulated upon transformation into yeast. We indeed...
Fork stabilization at DNA impediments is key to maintaining replication fork integrity and preventing chromosome breaks. Mrc1 Tof1 are two known stabilizers that travel with the fork. In addition a structural role, has damage checkpoint function. Using yeast model system, we analyzed role of expanded CAG repeats medium long lengths, which stall forks cause trinucleotide expansion diseases such as Huntington's disease myotonic dystrophy. We demonstrate stabilizer but not activation function...
Abstract Aims The transcription factor Tbx5 controls cardiogenesis and drives Scn5a expression in mice. We have identified two variants TBX5 encoding p. D111Y F206L Tbx5, respectively, unrelated patients with structurally normal hearts diagnosed long QT (LQTS) Brugada (BrS) syndrome. Here, we characterized the consequences of each variant to unravel underlying disease mechanisms. Methods results combined clinical analysis vivo vitro electrophysiological molecular techniques human-induced...
We systematically varied conditions of twodimensional (2D) agarose gel electrophoresis to optimize separation DNA topoisomers that differ either by the extent knotting, catenation or supercoiling.To this aim we compared electrophoretic behavior three different families topoisomers: (i) supercoiled molecules, where supercoiling covered range extending from covalently closed relaxed up naturally molecules; (ii) postreplicative catenanes with number increasing 1 ∼15, both catenated rings were...
Key features• Preparative method to enrich DNA samples of supercoiled, catenated, and knotted families topoisomers, later analyzed by 2D gels (or other techniques, e.g., microscopy).• facilitate the separation topoisomers any given circular molecule.• Separation molecules with same molecular masses but different shapes can be optimized modifying conditions gels.• Evaluating roles electric field agarose concentration on electrophoretic mobility sheds light their physical characteristics.
A novel rare mutation in the pore region of Nav1.5 channels (p.L889V) has been found three unrelated Spanish families that produces quite diverse phenotypic manifestations (Brugada syndrome, conduction disease, dilated cardiomyopathy, sinus node dysfunction, etc.) with variable penetrance among families. We clinically characterized carriers and recorded Na+ current (INa) generated by p.L889V native (WT) channels, alone or combination, to obtain further insight into genotypic–phenotypic...
The ZFHX3 and SCN5A genes encode the zinc finger homeobox 3 (Zfhx3) transcription factor (TF) human cardiac Na+ channel (Nav1.5), respectively. effects of Zfhx3 on expression Nav1.5 channel, in excitability, are currently unknown. Additionally, we identified three variants probands diagnosed with familial atrial fibrillation (p.M1260T) Brugada Syndrome (p.V949I p.Q2564R). Here, analyzed native (WT) mutated current (INa) recorded HL-1 cardiomyocytes. mRNA can be detected ventricular samples....
Abstract Synapse-Associated Protein 97 (SAP97) is an anchoring protein that in cardiomyocytes targets to the membrane and regulates Na + K channels. Here we compared electrophysiological effects of native (WT) p.P888L SAP97, a common polymorphism. Currents were recorded from mice trans-expressing human WT or SAP97 Chinese hamster ovary (CHO)-transfected cells. The duration action potentials QT interval significantly shorter p.P888L-SAP97 than WT-SAP97 mice. Compared WT, increased charge...
Abstract Large-scale expansion of (GAA)n repeats in the first intron FXN gene is responsible for severe neurodegenerative disease, Friedreich’s ataxia humans. We have previously conducted an unbiased genetic screen GAA repeat instability a yeast experimental system. The majority genes that came from this encoded components DNA replication machinery, strongly implying irregularities are at heart expansions. This screen, however, also produced two unexpected hits: members CST complex, CDC13...
ATP-sensitive potassium (KATP) channels composed of Kir6.x and sulfonylurea receptor (SURs) subunits couple cellular metabolism to electrical activity. Cantú syndrome (CS) is a rare disease caused by mutations in the genes encoding Kir6.1 (KCNJ8) SUR2A (ABCC9) that produce KATP channel hyperactivity due reduced block physiological ATP concentrations. We functionally characterized p.S1054Y mutation identified two CS carriers, who exhibited mild phenotype although was predicted as highly...
We have addressed the question of nature Moloney murine leukemia virus (MoMuLV) repression in mouse embryos by assaying for transient expression MoMuLV-derived constructs microinjected into early cleavage embryos. show that same cis-acting DNA sequences responsible block MoMuLV embryonal carcinoma cell lines operate embryos: (i) long terminal repeat is nonfunctional, and (ii) +147 to +163 repressor binding site, or negative regulatory element, negatively regulates from an active promoter.
DNA topoisomerases are thought to play a critical role in transcription, replication and recombination as well the condensation segregation of sister duplexes during cell division. Here, we used high-resolution two-dimensional agarose gel electrophoresis study intermediates final products small circular linear minichromosomes Saccharomyces cerevisiae presence absence topoisomerase 2. The results obtained confirmed that whereas for minichromosomes, catenated accumulated 2, YACs were able...
DNA knots are deleterious for living cells if not removed. Several theoretical and simulation approaches address the question of how topoisomerases select intermolecular passages that preferentially lead to unknotting rather than knotting randomly fluctuating molecules, but formation in vivo remains poorly understood. form non-replicating replicating supercoiling as well intertwining thought play a crucial role both resolution by topoisomerase IV. To confirm this idea, we used...
DNA topoisomerases are the enzymes that regulate topology in all living cells. Since discovery and purification of ω (omega), when first were topoisomerase identified, function many has been examined. However, their ability to relax supercoiling unlink pre-catenanes partially replicated molecules received little attention. Here, we used two-dimensional agarose gel electrophoresis test three type II vitro: prokaryotic gyrase, IV human 2α. We examined proficiency these on a bacterial plasmid:...
1 Abstract Human disease Friedreich’s ataxia (FRDA) is caused by large-scale expansions of (GAA)n repeats in the first intron FXN gene. While repeat during intergenerational transmissions are causative for development, somatic additionally contribute to progression. We and others have previously shown that transiently pause replication fork progression cultured human cells. However, whether which mechanisms stalling underlies remained unclear. Here we developed a new genetically tractable...
Abstract The human disease Friedreich’s ataxia (FRDA) is caused by expansions of GAA•TTC repeats in the first intron frataxin ( FXN) gene, and both intergenerational somatic are crucial for development. We others have shown earlier that expanded can form an intramolecular triplex structure (H-DNA). Here we studied effects locked nucleic acid (LNA)-DNA mixmer oligonucleotides peptide (PNA) oligomers on expansion cultured cells. Our experimental system employes a mammalian/yeast shuttle...
ABSTRACT Background In a family with inappropriate sinus tachycardia (IST) we identified novel mutation (p.V240M) of the hyperpolarization-activated cyclic nucleotide-gated type 4 (HCN4) channel, which contributes to pacemaker current (I f ) in human sinoatrial node cells. Here clinically study and functionally analyze p.V240M variant. Methods Macroscopic HCN4 single-channel currents were recorded using patch-clamp cells expressing native (WT) and/or channels. Results All carriers exhibited...
Abstract Background Zfhx3 (zinc finger homeobox 3) is a transcription factor (TF) encoded by the ZFHX3 gene. GWAS and gene-based association studies showed that one of major atrial fibrillation (AF) susceptibility-conferring genes. The sodium current (INa), carried Nav1.5 channels SCN5A, responsible for ventricular action potential depolarization determines cardiac excitability. interacts with other TFs, such as Tbx5 Pitx2c increase decrease INa, respectively. However, effects on INa are...