A5077867147- Alzheimer's disease research and treatments
- HIV Research and Treatment
- HIV/AIDS drug development and treatment
- Neuroscience and Neuropharmacology Research
- Advanced Fluorescence Microscopy Techniques
- Cellular transport and secretion
- Microtubule and mitosis dynamics
- Advanced Biosensing Techniques and Applications
- Parkinson's Disease Mechanisms and Treatments
- Photosynthetic Processes and Mechanisms
- HIV-related health complications and treatments
- Mitochondrial Function and Pathology
- DNA and Nucleic Acid Chemistry
- bioluminescence and chemiluminescence research
- Prion Diseases and Protein Misfolding
- DNA Repair Mechanisms
- Bioinformatics and Genomic Networks
- Genomics and Chromatin Dynamics
- Photoreceptor and optogenetics research
- Cytomegalovirus and herpesvirus research
- Enzyme Structure and Function
- BRCA gene mutations in cancer
- Gene expression and cancer classification
- Photochemistry and Electron Transfer Studies
- Receptor Mechanisms and Signaling
Scuola Normale Superiore
2011-2020
Istituto di Fisiologia Clinica
2013
University of Milano-Bicocca
2008-2010
Recent advances in fluorescence microscopy provided tools for the investigation and analysis of viral replication steps cellular context. In HIV field, current visualization systems successfully achieve fluorescent labeling envelope proteins, but not genome. Here, we developed a system able to visualize proviral DNA HIV-1 through immunofluorescence detection repair foci double-strand breaks specifically induced genome by heterologous expression I-SceI endonuclease. The Single-Cell Imaging...
Tau displacement from microtubules is the first step in onset of tauopathies and followed by toxic protein aggregation. However, other non-canonical functions might have a role these pathologies. Here, we demonstrate that small amount localizes nuclear compartment accumulates both soluble chromatin-bound fractions. We show favoring translocation accumulation, overexpression or detachment MTs, increases expression VGluT1, disease-relevant gene directly involved glutamatergic synaptic...
The ability to visualize fluorescent HIV-1 particles within the nuclei of infected cells represents an attractive tool study nuclear biology virus. To this aim we recently developed a microscopy-based system (HIV-IN-EGFP) that has proven valid efficiently complexes in compartment and examine import efficiency power method investigate viral events occurring between cytoplasmic is further shown through analysis HIV-IN-EGFP expressing TRIMCyp restriction factor. In these are not detected...
Formation of Tau aggregates is a common pathological feature tauopathies and their accumulation directly correlates with cytotoxicity neuronal degeneration. Great efforts have been made to understand aggregation find therapeutics halting or reversing the process, however, progress has slowed due lack suitable method for monitoring aggregation. We developed cell-based assay allowing detect quantify in living cells. The system based on FRET biosensor CST able monitor molecular dynamic...
The microtubule (MT)-associated protein Tau is a natively unfolded protein, involved in number of neurodegenerative disorders, collectively called tauopathies, aggregating neurofibrillary tangles (NFT). It an open question how the conversion from MT bound molecule to aggregation-prone species occurs and, also, if and tauopathy-related mutations affect its behavior cell. To address these points, we exploited genetically encoded FRET sensor based on full length monitor real time conformational...
The tumour-suppressor gene BRCA2 has been demonstrated to be involved in maintenance of genome integrity by affecting DNA double-strand break repair and homologous recombination. Protein-truncating mutations predispose women early onset breast ovarian cancers account for 15–30% familial cancer risk. In contrast, the human risk due missense mutations, intronic variants, in-frame deletions insertions gene, called unclassified not determined. Here, we want define if yeast Saccharomyces...
GFP mutants are known to display fluorescence flickering, a process that occurs in wide time range. Because serine 65, threonine 203, glutamate 222, and histidine 148 have been indicated as key residues determining the photodynamics, we focused here on role of 222 by studying dynamics GFPmut2 (S65A, V68L, S72A GFP) its H148G (Mut2G) E222Q (Mut2Q) mutants. Two relaxation components found autocorrelation functions GFPmut2: 10−100 μs pH-dependent component 100−500 laser-power-dependent...
ABSTRACT Recent advances in fluorescence microscopy allow three-dimensional analysis of HIV-1 preintegration complexes the nuclei infected cells. To extend this investigation to gammaretroviruses, we engineered a fluorescent Moloney murine leukemia virus (MLV) system consisting MLV-integrase fused enhanced green protein (MLV-IN-EGFP). A comparative lentiviral (HIV-1) and gammaretroviral cells revealed their different spatial distributions. This research tool has potential achieve new insight...
Endoplasmic reticulum (ER) dysfunction is important for alpha-synuclein (αS) acquired toxicity. When targeted to the ER in SH-SY5Y cells, transient or stable expression of αS resulted formation compact αS-positive structures a small subpopulation resembling inclusions. Thus, because limitations immunofluorescence, we developed set FRET biosensors (AFBs) able track conformation cells. In native conditions, i36, cell line αS, intermolecular AFBs, reporters which CFP YFP has been fused with...
New probes for kinetic intracellular measurements in the millisecond range are desirable to monitor protein biochemical dynamics essential catalysis, allosteric regulation, and signaling. Good candidates this aim photoswitchable mutants of green fluorescent protein, whose anionic fluorescence, primed by blue light, is markedly enhanced under an additional excitation at a shorter wavelength relaxes within few milliseconds. The report study how brightness enhancement kinetics depends on...
Tauopathies are neurodegenerative disorders characterized by Tau aggregation. Genetic studies on familial cases allowed for the discovery of mutations in MAPT gene that increase propensity to detach from microtubules and form insoluble cytoplasmic aggregates. Recently, rare mutation Q336H has been identified be associated with Pick’s disease (PiD) biochemical analyses demonstrated its ability (MTs) polymerization, thus revealing an opposite character compared other studied so far. Here we...
Tau is a microtubule binding protein expressed in neurons and its main known function related to the maintenance of cytoskeletal stability. However, recent evidence indicated that present also other subcellular compartments including nucleus where it implicated DNA protection, rRNA transcription, mobility retrotransposons structural organization nucleolus. We have recently demonstrated nuclear involved expression VGluT1 gene, suggesting molecular mechanism could explain pathological increase...
ABSTRACT Impaired interactions of Tau protein with microtubules (MT) and misfolding play a key role in Alzheimer disease (AD) other neurodegenerative diseases collectively named Tauopathies. However, little is known about the molecular conformational changes that underlie aggregation pathological conditions, due to difficulty studying structural aspects this intrinsically unfolded protein, particularly context living cells. Here we developed new Conformational-Sensitive sensor (CST), based...
Background Integration of HIV-1 provirus occurs mainly in transcriptionally active units. Furthermore PICs preferentially localize decondensed chromatin regions near the nuclear rim [1]. A technique recently developed our lab, Single Cell Imaging Provirus (SCIP), allowed us to determine that proviral DNA also localizes [2]. Emerging evidence suggests functions such as transcription, repair and replication are coordinated by a specific dynamic architecture not just linear sequence...
Background To study the nuclear biology of HIV-1 we recently developed a microscopy based fluorescent HIV-INEGFP system [1]. HIV-IN-EGFP exploits Vpr mediated trans-incorporation to incorporate IN-EGFP in viral particles made with pNL-IN-D64E. The ability visualize within nucleus makes it an attractive tool quantitatively import step [2,3]. In this work provide new evidence for efficient visualization complexes nuclei infected cells through optimized system.
Summary Tau displacement from microtubules is the first step in onset of tauopathies, and followed by toxic protein aggregation. However, other non-canonical functions might have a role these pathologies. Here, we demonstrate that small amount localizes nuclear compartment accumulates both soluble DNA-bound fractions. We show regulates expression VGluT1, disease-relevant gene directly involved glutamatergic synaptic transmission. Impeding Tau/tubulin interaction cytosol favours its...
Tau is a microtubule binding protein expressed in neurons and its main known function related to the maintenance of cytoskeletal stability. However, recent evidence indicated that present also other subcellular compartments including nucleus where it implicated DNA protection, rRNA transcription, mobility retrotransposons structural organization nucleolus. We have recently demonstrated nuclear involved expression VGluT1 gene, suggesting molecular mechanism could explain pathological increase...