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Screening Readthrough Compounds to Suppress Nonsense Mutations: Possible Application to β-Thalassemia

Monica Borgatti, Emiliano Altamura, Francesca Salvatori, Elisabetta D'Aversa, Nicola Altamura
2020 Journal of Clinical Medicine  
Several types of thalassemia (including β039-thalassemia) are caused by nonsense mutations in genes controlling globin production, leading to premature translation termination and mRNA destabilization mediated by the nonsense mediated mRNA decay. Drugs (for instance, aminoglycosides) can be designed to suppress premature translation termination by inducing readthrough (or nonsense suppression) at the premature termination codon. These findings have introduced new hopes for the development of a
more » ... harmacologic approach to cure this genetic disease. In the present review, we first summarize the principle and current status of the chemical relief for the expression of functional proteins from genes otherwise unfruitful for the presence of nonsense mutations. Second, we compare data available on readthrough molecules for β0-thalassemia. The examples reported in the review strongly suggest that ribosomal readthrough should be considered as a therapeutic approach for the treatment of β0-thalassemia caused by nonsense mutations. Concluding, the discovery of molecules, exhibiting the property of inducing β-globin, such as readthrough compounds, is of great interest and represents a hope for several patients, whose survival will depend on the possible use of drugs rendering blood transfusion and chelation therapy unnecessary.
doi:10.3390/jcm9020289 pmid:31972957 pmcid:PMC7073686 fatcat:vdefgwpyuzb73k5j4rrhk22gta

Recent Theoretical Approaches to Minimal Artificial Cells

Fabio Mavelli, Emiliano Altamura, Luigi Cassidei, Pasquale Stano
2014 Entropy  
Author Contributions Mavelli is the lead author of this line of research who has collaborated with Stano and Cassidei for specific topics, while Altamura is a young Ph.D. student who joined the group recently  ... 
doi:10.3390/e16052488 fatcat:dlvw5nartzdnnczhmboujaarji

The Origin and Early Evolution of Life: (Prebiotic) Systems Chemistry Perspective

Emiliano Altamura, Michele Fiore
2022 Life  
Acknowledgments: Michele Fiore and Emiliano Altamura wish to thank all the contributors of the Special Issue of Life (ISSN 2075-1729): "The Origin and Early Evolution of Life: Prebiotic Systems Chemistry  ... 
doi:10.3390/life12050710 pmid:35629377 pmcid:PMC9145544 fatcat:prgfm6ix7jez5n43to6yqswfva

Table-top combined scanning X-ray small angle scattering and transmission microscopies of lipid vesicles dispersed in free-standing gel

Francesco Scattarella, Emiliano Altamura, Paola Albanese, Dritan Siliqi, Massimo Ladisa, Fabio Mavelli, Cinzia Giannini, Davide Altamura
2021 RSC Advances  
X-ray microscopy combining absorption and SAXS contrast allows the graphical and quantitative description of spatial and concentration distribution of lipid vesicles, evidencing their interaction and aggregation, both in a wet and dry free-standing gel matrix.
doi:10.1039/d0ra08581b pmid:35423036 pmcid:PMC8690998 fatcat:mm5uc6uy2bclpe65mwuqsuopde

Chemical-Induced Read-Through at Premature Termination Codons Determined by a Rapid Dual-Fluorescence System Based on S. cerevisiae

Emiliano Altamura, Monica Borgatti, Alessia Finotti, Jessica Gasparello, Roberto Gambari, Mariangela Spinelli, Rosa Castaldo, Nicola Altamura, Georg Stoecklin
2016 PLoS ONE  
Nonsense mutations generate in-frame stop codons in mRNA leading to a premature arrest of translation. Functional consequences of premature termination codons (PTCs) include the synthesis of truncated proteins with loss of protein function causing severe inherited or acquired diseases. A therapeutic approach has been recently developed that is based on the use of chemical agents with the ability to suppress PTCs (read-through) restoring the synthesis of a functional full-length protein.
more » ... interest for compounds able to induce read-through requires an efficient high throughput large scale screening system. We present a rapid, sensitive and quantitative method based on a dual-fluorescence reporter expressed in the yeast Saccharomyces cerevisiae to monitor and quantitate read-through at PTCs. We have shown that our novel system works equally well in detecting read-through at all three PTCs UGA, UAG and UAA.
doi:10.1371/journal.pone.0154260 pmid:27119736 pmcid:PMC4847774 fatcat:co4gwzq4rbfudco4ajuwp2la4i

Racemic Phospholipids for Origin of Life Studies

Emiliano Altamura, Arnaud Comte, Alice D'Onofrio, Charlotte Roussillon, Dimitri Fayolle, René Buchet, Fabio Mavelli, Pasquale Stano, Michele Fiore, Peter Strazewski
2020 Symmetry  
Although prebiotic condensations of glycerol, phosphate and fatty acids produce phospholipid esters with a racemic backbone, most experimental studies on vesicles intended as protocell models have been carried out by employing commercial enantiopure phospholipids. Current experimental research on realistic protocell models urgently requires racemic phospholipids and efficient synthetic routes for their production. Here we propose three synthetic pathways starting from glycerol or from racemic
more » ... lketal (α,β-isopropylidene-dl-glycerol) for the gram-scale production (up to 4 g) of racemic phospholipid ester precursors. We describe and compare these synthetic pathways with literature data. Racemic phosphatidylcholines and phosphatidylethanolamines were obtained in good yields and high purity from 1,2-diacylglycerols. Racemic POPC (rac-POPC, (R,S)-1-palmitoyl-2-oleoyl-3-phosphocholine), was used as a model compound for the preparation of giant vesicles (GVs). Confocal laser scanning fluorescence microscopy was used to compare GVs prepared from enantiopure (R)-POPC), racemic POPC (rac-POPC) and a scalemic mixture (scal-POPC) of (R)-POPC enriched with rac-POPC. Vesicle morphology and size distribution were similar among the different (R)-POPC, rac-POPC and scal-POPC, while calcein entrapments in (R)-POPC and in scal-POPC were significantly distinct by about 10%.
doi:10.3390/sym12071108 fatcat:rjoj6mkypbe5zliwyvb5lmnbg4

Light-driven ATP production promotes mRNA biosynthesis inside hybrid multi-compartment artificial protocells [article]

Emiliano Altamura, Paola Albanese, Roberto Marotta, Francesco Milano, Michele Fiore, Massimo Trotta, Pasquale Stano, Fabio Mavelli
2020 bioRxiv   pre-print
Altamura, E., Carrara, P., D'Angelo, F., Mavelli, F. & Stano, P.  ... 
doi:10.1101/2020.02.05.933846 fatcat:efjcjgoa35hmhkujl5k64pyqkq

The Rise of the Nested Multicompartment Model in Synthetic Cell Research

Emiliano Altamura, Paola Albanese, Fabio Mavelli, Pasquale Stano
2021 Frontiers in Molecular Biosciences  
Image taken from Altamura et al. (2017) with the permission of the National Academy of Science United States ©2017.  ...  Alternatively, "prefabricated" and highly efficient organellae have been used, borrowing them from photosynthetic bacteria of the genus Rhodobacter (Altamura et al., 2021) (Figure 1G ).  ... 
doi:10.3389/fmolb.2021.750576 pmid:34540903 pmcid:PMC8446550 fatcat:hxmj6y5smnhaxj6zpctblgxkjy

Novel directions in molecular systems design: The case of light-transducing synthetic cells

Pasquale Stano, Emiliano Altamura, Fabio Mavelli
2017 Communicative & Integrative Biology  
Addendum to: Altamura, E.; Milano, F.; Tangorra, R. R.; Trotta, M.; Hassan Omar, O.; Stano, P.; Mavelli, F.  ...  program focused on the construction of photoautotrophic synthetic cells, starting with the reconstitution, in active and highly oriented form, of the photosynthetic reaction centre in giant lipid vesicles (Altamura  ... 
doi:10.1080/19420889.2017.1365993 pmid:29260799 pmcid:PMC5731512 fatcat:5mwfycanqnd5ddxkoq37jyo7fq

Light emitting biosilica by in vivo functionalization of Phaeodacty-lum tricornutum diatom microalgae with organometallic complexes [article]

Vona Danilo, Ragni Roberta, Altamura Emiliano, Albanese Paola, Giangregorio Michelaria, R. Cicco Stefania, M. Farinola Gianluca
2021 Zenodo  
Author Contributions: Danilo Vona contributed to diatoms culture and feeding experiments; Roberta Ragni contributed to complex synthesis and characterization, experimental design and paper writing; Emiliano  ...  Altamura and Paola Albanese performed confocal microscopy analysis; Michelaria Giangregorio contributed to Raman analysis; Stefania Roberta Cicco contributed to the spectroscopic characterization of biosilica  ... 
doi:10.5281/zenodo.5749365 fatcat:eahkomoojbapnls5xtvsoszcxq

Charge Recombination Kinetics of Bacterial Photosynthetic Reaction Centres Reconstituted in Liposomes: Deterministic Versus Stochastic Approach

Emiliano Altamura, Paola Albanese, Pasquale Stano, Massimo Trotta, Francesco Milano, Fabio Mavelli
2020 Data  
In this theoretical work, we analyse the kinetics of charge recombination reaction after a light excitation of the Reaction Centres extracted from the photosynthetic bacterium Rhodobacter sphaeroides and reconstituted in small unilamellar phospholipid vesicles. Due to the compartmentalized nature of liposomes, vesicles may exhibit a random distribution of both ubiquinone molecules and the Reaction Centre protein complexes that can produce significant differences on the local concentrations from
more » ... the average expected values. Moreover, since the amount of reacting species is very low in compartmentalized lipid systems the stochastic approach is more suitable to unveil deviations of the average time behaviour of vesicles from the deterministic time evolution.
doi:10.3390/data5020053 fatcat:wawznz4rivbohc7w55anhysnlm

Measurement and Numerical Modeling of Cell-Free Protein Synthesis: Combinatorial Block-Variants of the PURE System

Paolo Carrara, Emiliano Altamura, Francesca D'Angelo, Fabio Mavelli, Pasquale Stano
2018 Data  
Protein synthesis is at the core of bottom-up construction of artificial cellular mimics. Intriguingly, several reports have revealed that when a transcription–translation (TX–TL) kit is encapsulated inside lipid vesicles (or water-in-oil droplets), high between-vesicles diversity is observed in terms of protein synthesis rate and yield. Stochastic solute partition can be a major determinant of these observations. In order to verify that the variation of TX–TL components concentration brings
more » ... ut a variation of produced protein rate and yield, here we directly measure the performances of the 'PURE system' TX–TL kit variants. We report and share the kinetic traces of the enhanced Green Fluorescent Protein (eGFP) synthesis in bulk aqueous phase, for 27 combinatorial block-variants. The eGFP production is a sensitive function of TX–TL components concentration in the explored concentration range. Providing direct evidence that protein synthesis yield and rate actually mirror the TX–TL composition, this study supports the above-mentioned hypothesis on stochastic solute partition, without excluding, however, the contribution of other factors (e.g., inactivation of components).
doi:10.3390/data3040041 fatcat:5ut7cyi4svepjmp6ljdkv5q4pq

Self-division of giant vesicles driven by an internal enzymatic reaction

Ylenia Miele, Zsófia Medveczky, Gábor Holló, Borbála Tegze, Imre Derényi, Zoltán Hórvölgyi, Emiliano Altamura, István Lagzi, Federico Rossi
2020 Chemical Science  
Giant unilamellar vesicles having pH-sensitive bilayers can undergo self-division triggered by an internal enzymatic (urea–urease) reaction coupled to a cross-membrane transport of the chemical species.
doi:10.1039/c9sc05195c pmid:34122829 pmcid:PMC8157745 fatcat:ildlonajqbda5meoxg7ps3ocgu

Light-Emitting Biosilica by In Vivo Functionalization of Phaeodactylum tricornutum Diatom Microalgae with Organometallic Complexes

Danilo Vona, Roberta Ragni, Emiliano Altamura, Paola Albanese, Maria Michela Giangregorio, Stefania Roberta Cicco, Gianluca Maria Farinola
2021 Applied Sciences  
In vivo incorporation of a series of organometallic photoluminescent complexes in Phaeodactylum tricornutum diatom shells (frustules) is investigated as a biotechnological route to luminescent biosilica nanostructures. [Ir(ppy)2bpy]+[PF6]−, [(2,2′-bipyridine)bis(2-phenylpyridinato)iridium(III) hexafluorophosphate], [Ru(bpy)3]2+ 2[PF6]−, [tris(2,2′-bipyridine)ruthenium(II) hexafluorophosphate], AlQ3 (tris-(8-hydroxyquinoline)aluminum), and ZnQ2 (bis-8-hydroxyquinoline-zinc) are used as model
more » ... lexes to explore the potentiality and generality of the investigated process. The luminescent complexes are added to the diatom culture, and the resulting luminescent silica nanostructures are isolated by an acid-oxidative treatment that removes the organic cell matter without altering both frustule morphology and photoluminescence of incorporated emitters. Results show that, except for ZnQ2, the protocol successfully leads to the incorporation of complexes into the biosilica. The spontaneous self-adhering ability of both bare and doped Phaeodactylum tricornutum cells on conductive indium tin oxide (ITO)-coated glass slides is observed, which can be exploited to generate dielectric biofilms of living microorganisms with luminescent silica shells. In general, this protocol can be envisaged as a profitable route to new functional nanostructured materials for photonics, sensing, or biomedicine via in vivo chemical modification of diatom frustules with organometallic emitters.
doi:10.3390/app11083327 fatcat:semrx6wr75htlgl6iqub6nth5i

Incorporating a molecular antenna in diatom microalgae cells enhances photosynthesis [article]

Gabriella Leone, De La Cruz Valbuena Gabriel, R. Cicco Stefania, Danilo Vona, Altamura Emiliano, Ragni Roberta, Molotokaite Egle, Cecchin Michela, Cazzaniga Stefano, Ballottari Matteo, D'Andrea Cosimo, Lanzani Guglielmo (+1 others)
2021 Zenodo  
Diatom microalgae have great industrial potential as next-generation sources of biomaterials and biofuels. Effective scale-up of their production can be pursued by enhancing the efficiency of their photosynthetic process in a way that increases the solar-to-biomass conversion yield. A proof-of-concept demonstration is given of the possibility of enhancing the light absorption of algae and increasing their efficiency in photosynthesis by in vivo incorporation of an organic dye which acts as an
more » ... tenna and enhances cell growth and biomass without resorting to genetic modification. A molecular dye (Cy5) is incorporated in Thalassiosira weissflogii diatom cells by simply adding it to the culture medium and thus filling the orange gap that limits their absorption of sunlight. Cy5 enhances diatoms' photosynthetic oxygen production and cell density by 49% and 40%, respectively. Cy5 incorporation also increases by 12% the algal lipid free fatty acid (FFA) production versus the pristine cell culture, thus representing a suitable strategy to enhance biofuel generation from algal species. Time-resolved spectroscopy reveals Förster Resonance Energy Transfer (FRET) from Cy5 to algal chlorophyll. The present approach lays the basis for non-genetic tailoring of diatoms' spectral response to light harvesting, opening up new ways for their industrial valorization
doi:10.5281/zenodo.5749336 fatcat:jyihnegxlrg37ch36kavqfd4pu
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