A5050232933- Fungal and yeast genetics research
- Protein Kinase Regulation and GTPase Signaling
- DNA Repair Mechanisms
- Enzyme Structure and Function
- Endoplasmic Reticulum Stress and Disease
- CRISPR and Genetic Engineering
- Cellular transport and secretion
- Cancer therapeutics and mechanisms
- ATP Synthase and ATPases Research
- PI3K/AKT/mTOR signaling in cancer
- Microtubule and mitosis dynamics
- Photosynthetic Processes and Mechanisms
- Polyamine Metabolism and Applications
- Mitochondrial Function and Pathology
- Genomics and Chromatin Dynamics
- Melanoma and MAPK Pathways
- Plant Stress Responses and Tolerance
- Carcinogens and Genotoxicity Assessment
- Telomeres, Telomerase, and Senescence
- Fungal Biology and Applications
- Protein Structure and Dynamics
- Synthesis and biological activity
- RNA modifications and cancer
- DNA and Nucleic Acid Chemistry
- RNA Research and Splicing
University of Milano-Bicocca
2015-2024
University of Milan
1998-2011
Abstract The yeast Rif2 protein is known to inhibit Mre11 nuclease and the activation of Tel1 kinase through a short motif termed MIN, which binds Rad50 subunit simulates its ATPase activity in vitro. mechanism by restrains consequences this inhibition at DNA double-strand breaks (DSBs) are poorly understood. In study, we employed AlphaFold-Multimer modelling pinpoint validate interaction surface between MIN Rad50. We also engineered rif2-S6E mutation that enhances inhibitory effect...
Recent epidemiological studies have demonstrated that the consumption of healthy foods are particularly rich in polyphenols might reduce incidence cancer and neurodegenerative diseases. In particular, chlorogenic acids (CGAs) occur ubiquitously food represent most abundant human diet. A number beneficial biological effects CGAs, such as anti-inflammatory activity, anti-carcinogenic protection against diseases, been reported. However, molecular mechanisms at base these activities not yet...
Activation of the checkpoint protein Tel1 requires Mre11-Rad50-Xrs2 (MRX) complex, which recruits at DNA double-strand breaks (DSBs) through direct interaction between and Xrs2. However, in vitro activation by MRX ATP binding to Rad50, suggesting a role also for MR subcomplex activation. Here we describe two separation-of-functions alleles, mre11-S499P rad50-A78T, show specifically affect without impairing functions DSB repair. Both Mre11-S499P Rad50-A78T reduce Tel1-MRX leading poor...
Intracellular calcium is a second messenger involved in several processes yeast, such as mating, nutrient sensing, stress response and cell cycle events. It was reported that glucose addition stimulates rapid increase free level yeast. To investigate the variations induced by different stimuli we used reporter system based on photoprotein aequorin. Glucose (50 mM) to nutrient‐starved cells an intracellular concentration, mainly due influx from external medium. The of reached its maximum...
Though fungi show peculiarities in the purposes and specific traits of calcium signaling pathways, general scheme most important players are well conserved if compared to higher eukaryotes. This provides a powerful opportunity either investigate shared features using yeast as model or exploit fungal specificities potential targets for antifungal therapies. The sequenced genomes from <em>Saccharomyces cerevisiae, Schizosaccharomyces pombe</em> filamentous fungus <em>Neurospora crassa</em>...
The Mre11-Rad50-Xrs2 (MRX) complex detects and processes DNA double-strand breaks (DSBs). Its binding processing activities are regulated by transitions between an ATP-bound state a post-hydrolysis cutting that is nucleolytically active. Mre11 endonuclease activity stimulated Sae2, whose lack increases MRX persistence at DSBs checkpoint activation. Here we show the Rif2 protein inhibits responsible for increased retention in sae2Δ cells. We identify Rad50 residue important Rad50-Rif2...
Cannabidiol (CBD) is a non-psychoactive phytocannabinoid that has been discussed for its safety and efficacy in cancer treatments. For this reason, we have inquired into use on triple-negative human breast cancer. Analyzing the biological effects of CBD MDA-MB-231, demonstrated both dosage serum concentrations culture medium influence outcomes; furthermore, light scattering studies impacts aggregation state by acting as surfactant agent. Pharmacological combination with chemotherapeutic...
Abstract DNA double-strand breaks (DSBs) can be repaired by either homologous recombination (HR) or non-homologous end-joining (NHEJ). NHEJ is induced the binding to DSBs of Ku70–Ku80 heterodimer, which acts as a hub for recruitment downstream components. An important issue in DSB repair maintenance ends close proximity, function that yeast involves MRX complex and Sae2. Here, we provide evidence Ku contributes keep tethered each other. The ku70-C85Y mutation, increases affinity its...
Sae2 cooperates with the Mre11-Rad50-Xrs2 (MRX) complex to initiate resection of DNA double-strand breaks (DSBs) and maintain DSB ends in close proximity allow their repair. How these diverse MRX-Sae2 functions contribute damage resistance is not known. Here, we describe mre11 alleles that suppress hypersensitivity sae2Δ cells genotoxic agents. By assessing impact mutations at cellular structural levels, found all restore both camptothecin phleomycin affect Mre11 N-terminus defect by...
The plasma membrane H+-ATPase from Saccharomyces cerevisiae is an enzyme that plays a very important role in the yeast physiology. addition of protonophores, such as 2,4-dinitrophenol (DNP) and carbonyl cyanide m-chlorophenylhydrazone (CCCP), also triggers clear vivo activation this enzyme. Here, we demonstrate CCCP-induced shares some similarities with sugar-induced Phospholipase C protein kinase activities are essential for process while Gpa2p, G involved glucose-induced ATPase, not...
In Saccharomyces cerevisiae the Cdc25/Ras/cAMP pathway is involved in cell growth and proliferation regulation. Ras proteins are regulated by Ira1/2 GTPase activating (GAPs) Cdc25/Sdc25 guanine nucleotide exchange factors (GEFs). Most of cytosolic Cdc25 protein was found on internal membranes exponentially growing cells, while upon incubation a buffer with no nutrients it re-localized to plasma membrane. The overexpression Tpk1 PKA catalytic subunit also induces export from nucleus,...
This protocol describes the use of jellyfish Aequorea victoria aequorin protein to measure Ca 2+ levels in living yeast cells. All strains be analyzed must express A. apoprotein calcium biosensor, reconstituted into fully active by association with its cofactor, coelenterazine, which cannot synthesized itself. The simplest way achieve reconstitution is transform cells a vector driving apoaequorin expression, and then supply commercially available coelenterazine cofactor medium....