We studied the effect of branched-chain polyamines on the folding transition of genome-sized DNA molecules in aqueous solution by the use of single-molecule observation with fluorescence microcopy. Detailed morphological features of polyamine/DNA complexes were characterized by atomic force microscopy (AFM). The AFM observations indicated that branched-chain polyamines tend to induce a characteristic change in the higher-order structure of DNA by forming bridges or crosslinks between the segments of a DNA molecule. In contrast, natural linear-chain polyamines cause a parallel alignment between DNA segments. Circular dichroism measurements revealed that branched-chain polyamines induce the A-form in the secondary structure of DNA, while linear-chain polyamines have only a minimum effect. This large difference in the effects of branched- and linear-chain polyamines is discussed in relation to the difference in the manner of binding of these polyamines to negatively charged double-stranded DNA.

A versatile solid-phase approach based on peptide chemistry was used to construct four classes of structurally diverse polyamines with modified backbones: linear, partially constrained, branched, and cyclic. Their effects on DNA duplex stability and structure were examined. The polyamines showed distinct activities, thus highlighting the importance of polyamine backbone structure. Interestingly, the rank order of polyamine ability for DNA compaction was different to that for their effects on circular dichroism and melting temperature, thus indicating that these polyamines have distinct effects on secondary and higher-order structures of DNA.

Derivatives of the highly antitumor-active compound [{cis-Pt(NH)}(μ-OH)(μ-tetrazolato-N2,N3)] (5-H-Y), which is a tetrazolato-bridged dinuclear platinum(II) complex, were prepared by substituting a linear alkyl chain moiety at C5 of the tetrazolate ring. The general formula for the derivatives is [{cis-Pt(NH)}(μ-OH)(μ-5-R-tetrazolato-N2,N3)], where R is (CH)CH and n = 0 to 8 (complexes 1-9). The cytotoxicity of complexes 1-4 in NCI-H460 human non-small-cell lung cancer cells decreased with increasing alkyl chain length, and those of complexes 5-9 increased with increasing alkyl chain length. That is, the in vitro cytotoxicity of complexes 1-9 was found to have a U-shaped association with alkyl chain length. This U-shaped association is attributable to the degree of intracellular accumulation. Although circular dichroism spectroscopic measurement indicated that complexes 1-9 induced comparable conformational changes in the secondary structure of DNA, the tetrazolato-bridged complexes induced different degrees of DNA compaction as revealed by a single DNA measurement with fluorescence microsopy, which also had a U-shaped association with alkyl chain length that matched the association observed for cytotoxicity. Complexes 7-9, which had alkyl chains long enough to confer surfactant-like properties to the complex, induced DNA compaction 20 or 1000 times more efficiently than 5-H-Y or spermidine. A single DNA measurement with transmission electron microscopy revealed that complex 8 formed large spherical self-assembled structures that induced DNA compaction with extremely high efficiency. This result suggests that these structures may play a role in the DNA compaction that was induced by the complexes with the longer alkyl chains. The derivatization with a linear alkyl chain produced a series of complexes with unique cellular accumulation and DNA conformational change profiles and a potentially useful means of developing next-generation platinum-based anticancer drugs. In addition, the markedly high ability of these complexes to induce DNA compaction and their high intracellular accumulation emphasized the difference in mechanism of action from platinum-based anticancer drugs.

No abstract

The repetitive TG-rich DNA sequence at the chromosome end, telomere, protects cells from the end-replication problem in eukaryotic cells. S. cerevisiae telomerase (Est2) is the reverse transcriptase responsible for extending telomeres in yeast. Pif1 helicase has been implicated in regulating the telomerase activity. We used single-molecule experiments to investigate how Pif1 helicases regulate telomerase activity. We found that Est2 telomerase stayed bound to telomere end after extension, but Pif1 helicases remove telomerase from the telomere. In the presence of Pif1 helicases, multiple runs of the telomerase-mediated telomere lengthening were observed. This suggests a model that Pif1 helicases remove telomerase from the telomere ends, allowing telomerase recycling.

Recently, the influence of radiation on human body has been recognized as a serious problem. In particular, highly-reactive radicals produced by the radiation react with DNA, resulting in damage on its structure and electronic properties. It is thus important to investigate the reaction mechanism of radicals to DNA for elucidating the initial damage in DNA induced by the radiation. In the present study, we search for the transition states of the attacking mechanism between base-pair (G-C or A-T) and radical (OH and H radicals) in vacuum and in water, using the density functional theory calculations. The results elucidate that OH radical can cause mutation in A-T base pair, while H radical affects significantly on G-C base pair. 2P122アルコールによって誘起される DNA の凝縮・脱凝縮二段階 転移 Condensed DNA is unfolded into elongated conformation at ethanol concentration around 80%