The aim of this Special Issue is to highlight significant and new aspects concerning the chemistry and biology of noncanonical nucleic acid structures, with emphasis on their structure, stability, ...and conformational equilibria, as well as on the biological relevance of their interactions with proteins and ligands ....
The Lamiaceae family encompasses numerous species highly valued for their applications in medicine, food, and cosmetics. In order to screen the Lamiaceae family and discover new sources of ...phytochemicals and antioxidants, we comprehensively evaluated 20 species from this family, including Phlomis herba-venti, P. tuberosa, P. olivieri, P. kurdica, Nepeta sp., N. cataria, N. saccharata, Stachys sp., S. inflata, Scutellaria albida, Marrubium parviflora, Mentha pulegium, Thymus kotschyanus, Lamium album, Salvia officinalis, S. multicaulis, S. macrochlamys, S. candidissima, S. verticillata, and S. nemorosa. The aerial parts of these species were analyzed to determine their total phenolic (TPC) and flavonoid (TFC) contents, total tannin content (TTC), ascorbic acid content (AAC), antioxidant capacity (assessed by FRAP and DPPH assays), and polyphenolic components (by HPLC). The phytochemical compounds and antioxidant properties varied widely among different species. The highest concentrations of TPC (70.93 mg GAE/g DW), TFC (17.89 mg Que/g DW), TTC (6.49 mg TAE/100 g), and AAC (1.15 mg AA/g DW), as well as the greatest antioxidant activity, were observed in different Salvia species. Additionally, chlorogenic and rosmarinic acids were the primary phenolic compounds identified in the extracts from the investigated Lamiaceae family. According to Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA), three groups of species were identified, characterized by variations in phytochemical composition and antioxidant capacity. The results obtained can provide new natural sources of phytochemicals and antioxidant agents, particularly from Salvia species, for the advancement of new products in the food, agricultural, cosmetics and health industries.
Targeting G-quadruplex structures is currently viewed as a promising anticancer strategy. Searching for potent and selective G-quadruplex binders, here we describe a small series of new monohydrazone ...derivatives designed as analogues of a lead which was proved to stabilize G-quadruplex structures and increase R loop levels in human cancer cells. To investigate the G-quadruplex binding properties of the new molecules, in vitro biophysical studies were performed employing both telomeric and oncogene promoter G-quadruplex-forming sequences. The obtained results allowed the identification of a highly selective G-quadruplex ligand that, when studied in human cancer cells, proved to be able to stabilize both G-quadruplexes and R loops and showed a potent cell killing activity associated with the formation of micronuclei, a clear sign of genome instability.
This communication reports on a possible distinct role of HMGB1 protein. Biophysical studies revealed that HMGB1 binds and stabilizes the G-quadruplex of the KRAS promoter element that is responsible ...for most of the transcriptional activity. Biological data showed that inhibition of HMGB1 increases KRAS expression. These results suggest that HMGB1 could play a role in the gene transcriptional regulation via the functional recognition of the G-quadruplex.
To selectively target telomeric G-quadruplex (G4) DNA, monomeric and dimeric naphthalene diimides (NDIs) were investigated as binders of multimeric G4 structures able to discriminate duplex DNA. ...These NDIs were analysed by the affinity chromatography-based screening G4-CPG (G-quadruplex on Controlled Pore Glass), using the sequence dAGGG(TTAGGG)7 (tel46), folding into two consecutive G4s, as model of the human telomeric G4 multimer. In parallel, a telomeric G4 monomer (tel26) and a duplex structure (ds27) were used as controls. According to G4-CPG screening, NDI-5 proved to be the best ligand in terms of dimeric G4 vs. duplex DNA selectivity and was analysed by circular dichroism (CD), gel electrophoresis, isothermal titration calorimetry (ITC) and fluorescence spectroscopy in its interactions with tel46. NDI-5 strongly binds and stabilizes tel46 G4, favouring a hybrid folding in K+-containing buffer. Under these conditions, the binding process comprises a first event involving three molecules of NDI-5 and a second one in which other six molecules bind to the DNA. In a metal cation-free system, NDI-5 induces tel46 G4 folding, as indicated by CD and PAGE, favouring an antiparallel structuring. Docking simulations showed that NDI-5 can effectively bind to the pocket between two G4 units, representing a promising ligand for multimeric G4s.
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•A small library of monomeric and dimeric naphthalene diimides (NDIs) was synthesized.•The binding to multimeric G-quadruplex (G4) structures, discriminating duplex DNA, was studied.•The affinity chromatography-based screening G4-CPG identified NDI-5 as the best ligand.•The NDI-5/multimer G4 interactions were investigated by CD, SVD, fluorescence, PAGE.•Docking studies showed NDI-5 intercalates into the G4-G4 pocket of the multimeric DNA model.
G-Quadruplexes (G4s) are noncanonical nucleic acid structures involved in the regulation of several biological processes of many organisms. The rational design of G4-targeting molecules developed as ...potential anticancer and antiviral therapeutics is a complex problem intrinsically due to the structural polymorphism of these peculiar DNA structures. The aim of the present work is to show how Ultraviolet Resonance Raman (UVRR) spectroscopy can complement other techniques in providing valuable information about ligand/G4 interactions in solution. Here, the binding of BRACO-19 and Pyridostatin - two of the most potent ligands - to selected biologically relevant G4s was investigated by polarized UVRR scattering at 266 nm. The results give new insights into the binding mode of these ligands to G4s having different sequences and topologies by performing an accurate analysis of peaks assigned to specific groups and their changes upon binding. Indeed, the UVRR data not only show that BRACO-19 and Pyridostatin interact with different G4 sites, but also shed light on the ligand and G4 chemical groups really involved in the interaction. In addition, UVRR results complemented by circular dichroism data clearly indicate that the binding mode of a ligand can also depend on the conformation(s) of the target G4. Overall, these findings demonstrate the utility of using UVRR spectroscopy in the investigation of G4s and G4-ligand interactions in solution.
An increasingly comprehension of the folding intermediate states of DNA G-quadruplexes (G4s) is currently an important scientific challenge, especially for the human telomeric (h-tel) G4s-forming ...sequences, characterized by a highly polymorphic nature. Despite the G-triplex conformation was proposed as one of the possible folding intermediates for the antiparallel and hybrid h-tel G4s, for the parallel h-tel topology with an all-anti guanine orientation, a vertical strand-slippage involving the G-triplets was proposed in previous works through microseconds-long standard molecular dynamics simulations (MDs). Here, in order to get further insights into the vertical strand-slippage and the folding intermediate states of the parallel h-tel G4s, we have carried out a Well-Tempered Metadynamics simulation (WT-MetaD), which allowed us to retrieve an ensemble of six G4s having two/G-tetrad conformations derived by the G-triplets vertical slippage. The insights highlighted in this work are aimed at rationalizing the mechanistic characterisation of the parallel h-tel G4 folding process.
Target selectivity is one of the main challenges in the search for small molecules able to act as effective and non-toxic anticancer and/or antiviral drugs. To achieve this goal, handy, rapid and ...reliable High Throughput Screening methodologies are needed. We here describe a novel functionalization for the solid phase synthesis of oligonucleotides on Controlled Pore Glass, including a flexible hexaethylene glycol spacer linking the first nucleoside through the nucleobase via a covalent bond stable to the final deprotection step. This allowed us preparing fully deprotected oligonucleotides still covalently attached to their supports. In detail, on this support we performed both the on-line synthesis of different secondary structure-forming oligonucleotides and the affinity chromatography-based screenings of conformation-selective G-quadruplex ligands. By using a fluorescent core-extended naphthalene diimide with different emitting response upon binding to sequences folding into G-quadruplexes of different topologies, we have been able to discriminate not only G-quadruplex vs. duplex DNA structures, but also different G-quadruplex conformations on the glass beads by confocal microscopy.
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•A functionalized Controlled Pore Glass incorporating hexaethylene glycol was developed.•G-quadruplex-forming oligonucleotides were synthesized on this novel solid support.•These oligonucleotide-bound supports allowed affinity chromatography-based screenings.•Putative G-quadruplex ligands were analyzed by a rapid, easy spectrophotometric test.•Fluorescence confocal microscopy discriminated G-quadruplex vs. duplex DNA on CPG.
The i-motif is a biologically relevant non-canonical DNA structure formed by cytosine-rich sequences. Despite the importance of the factors affecting the formation/stability of such a structure, like ...pH, cation type and concentration, no systematic study that simultaneously analysed their effect on the i-motif in vitro has been carried out so far. Therefore, here we report a systematic study that aims to evaluate the effect of these factors, and their possible interaction, on the formation of an i-motif structure. Our results confirm that pH plays the main role in i-motif formation. However, we demonstrate that the effect of the cation concentration on the i-motif is strictly dependent on the pH, while no significant differences are observed among the investigated cation types.
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