Entrapping of fullerenes, nanotubes, and inorganic nanoparticles by a DNA-chitosan complex: a method for nanomaterials removal.

We report a protocol for entrapping of various water-dispersed nanomaterials: fullerenes, multiwall carbon nanotubes, quantum dots (semiconductor nanoparticles), and gold nanorods, into a DNA-chitosan complex. In contrast to small-size nanomaterial particles, the bulky DNA-chitosan interpolyelectrolyte complex incorporating the dispersed nanomaterials can be easily separated from aqueous media by centrifugation, filtration, or decantation. While the removal of nanoparticles by centrifugation is equally efficient for every type of nanoparticles and reaches 100%, the higher efficiency of the nanomaterials removal by other two methods is favored by larger size of nanoparticles. The application of this entrapping protocol for removal of nanomaterials from water is discussed.

Thiol-mediated anchoring of silver cations to DNA for construction of nanofibers on DNA scaffold.

The formation of metal-containing Ag-mercaptoethanol (-Ag-S(R)-)(n) complexes on DNA chain scaffold was studied by UV spectroscopy, zeta potential measurement, and fluorescence and transmission electron microscopies. Experimental results made clear the mechanism of DNA mineralization and compaction, according to which intercalation of silver cations into DNA scaffold and further formation of (-Ag-S(R)-)(n) oligomeric complexes on DNA induce efficient DNA chain compaction by terminal Ag(+) cations. By transmission electron microscopy the formation of fiber-like DNA-templated nanostructures was observed. DNA-Ag-thiol complexes are promising for DNA-templated engineering of hybrid 1D nanostructures with adjustable chemical functionalities by choosing appropriate thiol ligand.

Photochemical modulation of DNA conformation by organic dications.

A group of azobenzene derivatives containing two quaternary ammonium groups with various intercharge distances between them was synthesised and used to control photochemically the conformation of genomic DNA by switching the distance between cationic ammonium groups in the dications. It was found that isomerisation of either dication from the trans form to cis resulted in an increase in the dication's efficiency for DNA compaction; this is associated with a decrease in intercharge distance between ammonium groups and leads to a better match of the binder's cationic groups to adjacent phosphate groups of DNA. Ammonium dications have several important advantages over the photosensitive surfactant type of diazobenzene reported earlier: they can be used at significantly lower (>100-fold) concentrations than photosensitive surfactants, and DNA conformation control can be performed over a broader concentration range of dications. The influence of intercharge distance in photosensitive dications on photo-induced DNA binding discrimination is discussed, and the molecular mechanism is proposed.

A facile method for the assessment of DNA damage induced by UV-activated nanomaterials.

Fluorescent microscopy observation of gene-size DNA (T4 phage DNA or λ phage DNA) was used to assess DNA damage induced by UV irradiation in the presence of nanomaterials, such as QDs (quantum dots: CdSe/ZnS semiconductor nanoparticles), the water-soluble fullerene derivative C(60)(OH)(n) (n = 6-12) and titanium oxide nanoparticles of 25 nm in diameter. The magnitude of DNA damage could be simply evaluated based on the degree of shortening of the stretched DNA image. This method showed that DNA damage was amplified by the action of QDs under irradiation by C-band (λ(max) = 254 nm) or B-band (λ(max) = 303 nm) UV. Smaller QDs that emitted higher-energy fluorescence (λ = 565 nm) induced more severe damage than medium- and larger-size QDs that emitted longer-wavelength fluorescence (λ = 605 and 705 nm, respectively). The fullerene derivative and TiO(2) nanoparticles caused DNA damage even under irradiation by A-band UV (λ(max) = 365 nm) and showed more severe DNA damage than QDs under similar conditions.

DNA-assisted "double-templating" approach for the construction of hollow meshed inorganic nanoshells.

A "double-templating" approach was elaborated to produce meshed nanoshells made of semiconductor material in a two-step process. First, DNA adsorption is templated by spherical nanobeads, and second, DNA is mineralized by an inorganic material (CdS). Dissolution of the core beads leaves nanometer-size shells, the surface structure of which represents a mineralized network of DNAs. This method demonstrates the opportunity to metalize an arbitrary three-dimensional template by depositing a network of nanowires.

Thermo-switching of the conformation of genomic DNA in solutions of poly(N-isopropylacrylamide).

We used poly(N-isopropylacrylamide) (PNIPAM) to control the conformation of genomic DNA by changing the temperature of a reaction solution and studied the DNA transition at the level of single DNA molecules. With this method, the conformation of long genomic DNA can be readily and reversibly switched between a very compact condensate and an unfolded macromolecule.

Conformational behavior of giant DNA through binding with Ag+ and metallization.

The conformational behavior of a long single-chain double-stranded DNA in solutions of free silver ions and silver nanoparticles generated via the reduction of AgNO3 by NaBH4 was monitored by fluorescence and electron microscopies and UV spectroscopy. The interaction of monovalent silver ions with DNA induces shrinking of a DNA-coiled polymer chain as a result of a decrease in the DNA persistence length through the complexation of Ag+ with DNA bases. In contrast, the reduction of silver ions by NaBH4 in DNA solutions triggers DNA compaction: a DNA transition from elongated coil state into a compact state. This transition is continuous, unlike the all-or-none discrete DNA compaction that is commonly seen with multications. It is suggested that the collapse of DNA is accompanied by growth aggregation of silver nanoparticles generated on the DNA template.

ATP-induced shrinkage of DNA with MukB protein and the MukBEF complex of Escherichia coli.

Fluorescence microscopic observation of individual T4 DNA molecules revealed that the MukBEF complex (bacterial condensin) and its subunit, the MukB (a member of the SMC [structural maintenance of chromosomes] superfamily) homodimer, of Escherichia coli markedly shrunk large DNA molecules in the presence of hydrolyzable ATP. In contrast, in the presence of ADP or ATP-gammaS, the conformation of DNA was almost not changed. This suggests that the ATPase activity of subunit MukB is essential for shrinking large DNA molecules. Stretching experiments on the shrunken DNA molecules in the presence of ATP and MukBEF indicated a cross-bridging interaction between DNA molecules.

Visualization of different pathways of DNA release from interpolyelectrolyte complex.

The release of double-stranded DNA from its interpolyelectrolyte complex with positively charged poly(allylamine hydrochloride) via exchange reaction with added polyanion, poly(sodium styrenesulfonate), is directly observed by fluorescence microscopy. It is shown that the pathways of DNA release depend essentially on the amount of added low-molecular-weight salt. At low salt content, the DNA release proceeds via the formation of an intermediate "beads-on-string" structure, whereas at high salt content the release goes directly from globule to coil states without any intermediate structures. The reasons for different character of DNA release are discussed.

Single-chain compaction of long duplex DNA by cationic nanoparticles: modes of interaction and comparison with chromatin.

The compaction of long duplex DNA by cationic nanoparticles (NP) used as a primary model of histone core particles has been investigated. We have systematically studied the effect of salt concentration, particle size, and particle charge by means of single-molecule observations-fluorescence microscopy (FM) and transmission electron microscopy (TEM)-and molecular dynamics (MD) simulations. We have found that the large-scale DNA compaction is progressive and proceeds through the formation of beads-on-a-string structures of various morphologies. The DNA adsorbed amount per particle depends weakly on NP concentration but increases significantly with an increase in particle size and is optimal at an intermediate salt concentration. Three different complexation mechanisms have been identified depending on the correlation between DNA and NPs in terms of geometry, chain rigidity, and electrostatic interactions: free DNA adsorption onto NP surface, DNA wrapping around NP, and NP collection on DNA chain.

Transcription of giant DNA complexed with cationic nanoparticles as a simple model of chromatin.

We prepared complexes of giant double-stranded DNA with cationic nanoparticles of 10-40 nm in diameter as an artificial model of chromatin and characterized the properties of changes in their higher-order conformation. We measured the changes in transcriptional activity that accompanied the DNA conformational transitions. Complete inhibition was found at excess concentrations of nanoparticles. In contrast, at intermediate stages of DNA binding with nanoparticles, the transcription activity of DNA survived, and this strongly depended on the size of the nanoparticles. For large nanoparticles of 40 nm, a decrease in transcriptional activity can be caused by the addition of only a small amount of nanoparticles. On the other hand, there was almost no inhibition of DNA transcriptional activity with the addition of small nanoparticles (10 nm) until very high concentrations, even under conditions that induced DNA compaction as revealed by single-DNA observation. At higher concentrations of 10-nm nanoparticles, DNA transcription activity decreased abruptly until it was completely inhibited. These results are discussed in relation to the actual size of the histone core, together with the mechanism of switching of transcriptional activity in eukaryotic cells.

Compaction of DNA on nanoscale three-dimensional templates.

There exist several important in vivo examples, where a DNA chain is compacted on interacting with nanoscale objects such as proteins, thereby forming complexes with a well defined molecular architecture. One of the well known manifestations of such a natural organization of a semi-flexible DNA chain on nanoscale objects is hierarchical DNA molecule assembly into a chromosome, which is mediated by cationic histone proteins at the first stages of compaction. The biological importance of this and other natural nanostructural organizations of the DNA molecule has inspired many theoretical and numerical studies to gain physical insight into this problem. On the other hand, the experimental model systems containing DNA and nanoobjects, which are important to extend our knowledge beyond natural systems, were almost unavailable until the last decade. Accelerating progress in nanoscale chemistry and materials science has brought about various nanoscale three-dimensional structures such as dendrimers, nanoparticles, and nanotubes, and thus has provided a basis for the next important step in creating novel DNA-containing nanostructures, modelling of natural DNA compaction, and verification of accumulated theoretical predictions on the interaction between DNA and nanoscale templates. This review is written to highlight this early stage of nano-inspired progress and it is focused on physico-chemical and biophysical experimental investigations as well as theoretical and numerical studies dedicated to the compaction of DNA on nanoscale three-dimensional templates.

Compaction of single-chain DNA by histone-inspired nanoparticles.

We elaborated a versatile experimental model of chromatin which consists of a single chain of long duplex DNA that interacts with well-defined cationic nanoparticles of various sizes. We found that the DNA compaction by nanoparticles is stepwise and progressive at the single-chain level. It is controlled by the ability of DNA to wrap nanoparticles, which is more efficient for larger particles and, similar to DNA-histone interaction, is optimal at a physiological salt concentration.

Specific formation of beads-on-a-chain structures on giant DNA using a designed polyamine derivative.

Fluorescence microscopy was used to study the folding transition of giant DNAs, T4 DNA (ca. 166 kbp), and lambda DNA (ca. 48 kbp), which proceeds through intermediates with intramolecular segregation induced by pteridine-polyamine conjugates, i.e., 2-amino-6,7-dimethyl-4-(4,9,13-triazatridecylamino)pteridine and -4-(3-(aminopropyl)amino)pteridine. According to the results of DNA denaturation, UV and fluorescent spectroscopy, and transmission electron microscopic observations, it became clear that DNA folding induced by the polyamine derivative is not a continuous shrinking process but a combination of discontinuous processes.

How does DNA compaction favor chiral selectivity with cationic species? Higher selectivity with lower cationic charge.

A single-molecule study on giant DNA compaction by enantiomeric dications and tetracations demonstrates that strong chiral discrimination in DNA compaction is manifested only if the positive charge on enantiomeric multications is relatively low. The increase in cationicity of the chiral compaction agent inevitably leads to an increase in nonspecific electrostatic interactions and quenching of chiral discrimination in the DNA-folding phase transition.

Na+ shows a markedly higher potential than K+ in DNA compaction in a crowded environment.

Whereas many physicochemical investigations have shown that among monovalent cations Na(+) ion possesses minimal potential for DNA binding, biological assays have shown that Na(+) ion (in contrast to K(+) ion) plays a primary role in chromatin compaction and related processes. It is difficult to explain this inverse relationship between the compaction potentials of Na(+) and K(+) and their binding abilities. In this study we sought to resolve this contradiction and emphasize the phenomenological distinction between DNA compaction and DNA binding processes in the case of DNA compaction by monocations. Using polyethylene glycol solutions as a model of a crowded cell environment, we studied DNA compaction by alkali metal salts LiCl, NaCl, KCl, RbCl, and CsCl, and found that all of these monocations promote DNA compaction. Among these monovalent cations Na(+) produces the greatest compaction and the ratio of K(+) cand Na(+) oncentrations for DNA compaction is approximately 1.5-2. A comparative analysis of recent experimental results indicates that a higher binding activity of monocation generally corresponds to a low compaction potential of the corresponding monovalent ion. This inverse relation is explained as a result of partial dehydration of monocations in the compact state.

DNA compaction by divalent cations: structural specificity revealed by the potentiality of designed quaternary diammonium salts.

DNA interaction with quaternary diammonium dications, R(CH(3))(2)N(+)(CH(2))(n)N(+)(CH(3))(2)R, having various intercharge distances, lengths, and branching, and the chemical nature of the hydrophobic substituents were investigated by fluorescent microscopy and circular dichroism (CD) spectroscopy to reveal their structural specificity for binding to DNA. The conformational behavior of DNA was found to be highly sensitive to the structure of the dications with separated charges. The distance between two ammonium groups greatly influences the compaction activity of the dications. To explain this situation, we proposed a model that demonstrates that the charge density of the dication and the geometric fit between DNA phosphates and the ammonium groups in the dications play an important role in providing efficient DNA collapse. Elongation of the alkyl substituents (R) in the diammonium salts from ethyl to hexyl did not generate any significant alterations in the compaction activities, whereas the branching of substituents caused a drastic decrease in their compaction ability. Based on the results of CD spectroscopy, it was found that the ability of the dications to provoke a DNA transition from the B-form to A-form was also specific: it depended on their intercharge distances and was independent of the length of alkyl substituents.

Ascorbic acid induces a marked conformational change in long duplex DNA.

Ascorbic acid is often regarded as an antioxidant in vivo, where it protects against cancer by scavenging DNA-damaging reactive oxygen species. However, the detailed mechanism of the action of ascorbic acid on genetic DNA is still unclear. We examined the effect of ascorbic acid on the higher-order structure of DNA through real-time observation by fluorescence microscopy. We found that ascorbic acid generates a pearling structure in single giant DNA molecules, with elongated and compact regions coexisting along a molecular chain. Results from electron microscopy and atomic force microscopy indicate that the compact regions assume a loosely packed conformation. A possible mechanism for the induction of this conformational change is discussed in relation to the interplay between the higher-order and second-order structures of DNA.