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Hidetaka Torigoe

Publications and source records attributed to Hidetaka Torigoe.

At least 19 recordsLinked to original sources
PubMed2006

Tetraplex structure of fission yeast telomeric DNA and unfolding of the tetraplex on the interaction with telomeric DNA binding protein Pot1.

To understand the regulation mechanism of fission yeast telomeric DNA, we analysed the structural properties of Gn: d(GnTTAC) (n=2-6) and 4Gn: d(GnTTAC)4 (n=3 and 4), and their interaction with the single-stranded telomeric DNA binding domain of telomere-binding protein Pot1 (Pot1DBD). G4, G5 and G6 formed a parallel tetraplex in contrast with no tetraplex formation by G2 and G3. Also, 4G4 adopted only an antiparallel tetraplex in spite of a mixture of parallel and antiparallel tetraplexes of 4G3. The variety of tetraplex structures was governed by the number of consecutive guanines in a single copy and the number of repeats. The antiparallel tetraplex of 4G4 became unfolded upon the interaction with Pot1DBD. The interaction with mutant Pot1DBD proteins revealed that the ability to unfold the antiparallel tetraplex was strongly correlated with the specific binding affinity for the single-stranded telomeric DNA. The result suggests that the decrease in the free single strand upon the complex formation with Pot1DBD may shift the equilibrium from the tetraplex to the single strand, which may cause the tetraplex unfolding. Considering that the antiparallel tetraplex inhibits telomerase-mediated telomere elongation, we conclude that the ability of Pot1 to unfold the antiparallel tetraplex is required for telomerase-mediated telomere regulation.

Circular Dichroism↗
PubMed2006

Detection of C:C mismatch base pair by fluorescence spectral change upon addition of silver (I) cation: toward the efficient analyses of single nucleotide polymorphism.

We have already found that silver (I) cation specifically binds to C:C mismatch base pair in heteroduplex, which increases the melting temperature of heteroduplex involving C:C mismatch base pair by about 4 degrees C. This result shows that addition of the metal cation is a promising strategy for the mismatch base pair detection in heteroduplex, but UV melting to determine the melting temperature is time-consuming. In the present study, to develop a more convenient way for the mismatch base pair detection, we examined the fluorescence spectral change of fluorescent-labelled duplex upon addition of the metal cation. Addition of silver (I) cation to the heteroduplex involving the C:C mismatch base pair significantly changed the fluorescence intensity, but no significant change in the fluorescence intensity was observed for the duplexes involving other base pairs. Our results certainly support the idea that the fluorescence spectral change upon the addition of the metal cation could be a convenient strategy for the mismatch base pair detection by the heteroduplex analysis, and may eventually lead to progress in SNP genotyping.

Base Pair Mismatch↗
PubMed2006

Detection of T:T mismatch base pair by fluorescence spectral change upon addition of mercury (II) cation: toward the efficient analyses of single nucleotide polymorphism.

We have already found that mercury (II) cation specifically binds to T:T mismatch base pair in heteroduplex, which increases the melting temperature of heteroduplex involving T:T mismatch base pair by about 4 degrees C. This result shows that addition of the metal cation is a promising strategy for the mismatch base pair detection in heteroduplex, but UV melting to determine the melting temperature is time-consuming. In the present study, to develop a more convenient way for the mismatch base pair detection, we examined the fluorescence spectral change of fluorescent-labelled duplex upon addition of the metal cation. Addition of mercury (II) cation to the heteroduplex involving the T:T mismatch base pair significantly changed the fluorescence intensity, but no significant change in the fluorescence intensity was observed for the duplexes involving other base pairs. Our results certainly support the idea that the fluorescence spectral change upon the addition of the metal cation could be a convenient strategy for the mismatch base pair detection by the heteroduplex analysis, and may eventually lead to progress in SNP genotyping.

Base Pair Mismatch↗
PubMed2006

Thermodynamic analyses of the specific interaction between two C:C mismatch base pairs and silver (I) cations.

We have already found that a single silver (I) cation specifically binds to a single C:C mismatch base pair in heteroduplex, which increases the melting temperature (T(m)) of heteroduplex involving a single C:C mismatch base pair by about 4 degrees C. Here, to examine the properties of the interaction between two C:C mismatch base pairs and silver (I) cations, we analyzed the effect of silver (I) cations on the thermal stability of heteroduplexes involving two C:C mismatch base pairs. The positions of the two C:C mismatch base pairs in heteroduplex did not significantly affect the magnitude of the increase in T(m) upon addition of silver (I) cation, suggesting that the binding affinity of the first silver (I) cation with one of the two C:C mismatch base pairs and that of the second silver (I) cation with the other of the two mismatch base pairs may be independent of the positions of the two mismatch base pairs. The larger magnitude of the increase in T(m) for the second silver (I) cation binding than that observed for the first silver (I) cation binding suggests that the binding affinity for the second silver (I) cation may be significantly higher than that for the first silver (I) cation. Our results certainly support the idea that addition of the metal cation is a promising strategy for the mismatch base pair detection in the heteroduplex analysis and may eventually lead to progress in SNP genotyping.

Base Pair Mismatch↗
PubMed2005

Synergistic stabilization of nucleic acid assembly by oligo-N3'-->P5' phosphoramidate modification and additions of comb-type cationic copolymers.

Synergic stabilization of DNA triplexes by oligo-N3'-->P5' phosphoramidate (PN) modification and additions of comb-type cationic copolymers was demonstrated. The combination of the copolymer and the PN modification increased triplex K(a) about 4 orders of magnitude. Kinetic analysis revealed that observed stabilization resulted from kinetic complimentarity between increased association rates by the copolymer and decreased dissociation rates by the PN modification of triplex forming oligonucleotides. No countering interference between these stabilizing effects was observed. We propose that kinetic analyses of stabilizing effects permit selection of a rational combination of stabilizing methods for successful synergy in stabilizing complex formation.

Amides↗
PubMed2005

Computational evaluation of the specific interaction between cation and mismatch base pair.

Ab initio calculations were carried out to characterize the structure and energetic of silver cation (Ag (I)) complex with cytosine (C:Ag:C) and mercury cation (Hg (II)) complex with thymine (T:Hg:T) systems. These metal-modified mismatch base pairs have been optimized using Hatree-Fock method without any symmetry constrains. Using above methods, the models of Ag (I) in a crosslink between O2 carbonyl oxygen of cytosine, O2(C):Ag:O2(C), and Hg (II) in a crosslink between N3 nitrogen atom of thymine, N3(T):Hg:N3(T) were obtained. Furthermore, the interaction energies of C:Ag:C and T:Hg:T models were estimated. The result showed that the coordination silver (I) cation with cytosine is more stable than hydrate state.

Base Pair Mismatch↗
PubMed2005

Thermodynamic analyses of the specific interaction between C:C mismatch base pair and silver (I) cation: toward the efficient detection of single nucleotide polymorphism.

We examined the effect of silver (I) cation on the thermal stability of heteroduplex and homoduplex. Addition of silver (I) cation increased the melting temperature of heteroduplex containing C:C mismatch base pair by about 4 degrees C. The thermal stability of homoduplex and heteroduplexes containing other kinds of mismatch base pairs was not significantly changed by the addition of silver (I) cation. Isothermal titration calorimetric study demonstrated that silver (I) cation directly bound to C:C mismatch base pair in heteroduplex at a molar ratio of 1:1. The binding constant and the enthalpy change for the binding of silver (I) cation to C:C mismatch base pair was approximately 10(6) M(-1) and -3 kcal mol(-1), respectively. We conclude that silver (I) cation directly binds to C:C mismatch base pair in heteroduplex with high affinity and specificity. Our results certainly support the idea that the addition of silver (I) cation to C:C mismatch base pair in heteroduplex could be a convenient strategy for heteroduplex analysis and may eventually lead to progress in single nucleotide polymorphism genotyping.

Base Pair Mismatch↗
PubMed2005

Development of triplex formation-based artificial transcription factor to recognize any upstream sequence of target genes.

Triplex formation-based artificial transcription factor to recognize any upstream sequence of desired genes was developed to regulate expression of any target genes. The reporter beta-galactosidase activity in yeast was increased 1.5-2 times by the introduction of the artificial transcription factor. The novel factor may be a useful tool to regulate expression of any target genes and reveal unknown function of the target genes.

Alcohol Dehydrogenase↗
PubMed2005

Tetraplex structure of fission yeast telomeric DNA and its unfolding by the interaction with telomeric DNA binding protein Pot1.

We compared the structure of fission yeast telomeric DNA, 4G4: d(GGGGTTAC)4, and nontelomeric DNA, 4T4: d(TTTTTTAC)4, and examined their interaction with telomeric DNA binding domain of telomeric DNA binding protein Pot1 (Pot1DBD). 4T4 did not form any higher-order structure, but 4G4 formed intramolecular folded tetraplex structure in the presence of Na+. Although Pot1DBD did not induce any significant structural change of 4T4, the intramolecular folded tetraplex structure of 4G4 was unfolded by the interaction with Pot1DBD. Pot1 may facilitate telomere elongation of telomerase by disrupting the tetraplex structure of the telomeric DNA.

DNA↗
PubMed2004

Silver (I) cation specifically stabilizes heteroduplex with C:C mismatch base pair: toward the efficient detection of single nucleotide polymorphism (2).

We examined the effect of silver (I) cation on the thermal stability of heteroduplex and homoduplex. Addition of silver (I) cation increased the melting temperature of heteroduplex containing C:C mismatch base pair by about 3-4 degrees C. The thermal stability of homoduplex and heteroduplexes containing other kinds of mismatch base pairs was not significantly changed by the addition of silver (I) cation. We conclude that silver (I) cation specifically stabilizes heteroduplex containing C:C mismatch base pair. Our results certainly support the idea that the addition of silver (I) cation to C:C mismatch base pair in heteroduplex could be a convenient strategy for heteroduplex analysis and may eventually lead to progress in single nucleotide polymorphism genotyping.

Base Pair Mismatch↗
PubMed2004

Synergistic stabilization of triplex by combination of comb-type cationic copolymer and 2',4'-BNA.

We examined the effect of the combination of the two triplex-stabilizing factors, poly(L-lysine)-graft-dextran (PLL-g-Dex) copolymer and 2'-O,4'-C-methylene bridged nucleic acid (2',4'-BNA) backbone modification of triplex-forming oligonucleotide (TFO), on the pyrimidine motif triplex formation at neutral pH, a condition where pyrimidine motif triplexes are unstable. The combination of both stabilizing factors that was the triplex involving the 2',4'-BNA TFO in the presence of the copolymer synergistically increased the thermal stability of the pyrimidine motif triplex at neutral pH. The present results certainly support the idea that the combination of the stabilizing factors can be a key method for triplex stabilization and may lead to progress in therapeutic applications of the antigene strategy in vivo.

DNA↗
PubMed2004

Thermodynamic analyses of the specific interaction between T:T mismatch base pair and mercury (II) cation: toward the efficient detection of single nucleotide polymorphism (1).

We examined the effect of mercury (II) cation on the thermal stability of heteroduplex and homoduplex. Addition of mercury (II) cation increased the melting temperature of heteroduplex containing T:T mismatch base pair by about 4 degrees C. The thermal stability of homoduplex and heteroduplexes containing other kinds of mismatch base pairs was not significantly changed by the addition of mercury (II) cation. Isothermal titration calorimetric study demonstrated that mercury (II) cation directly bound to T:T mismatch base pair in hcteroduplex at a molar ratio of 1:1. The binding constant and the enthalpy change for the binding of mercury (II) cation to T:T mismatch base pair was approximately 10(6) M(-1) and -6 kcal mol(-1), respectively. We conclude that mercury (II) cation directly binds to T:T mismatch base pair in heteroduplex with high affinity and specificity. Our results certainly support the idea that the addition of mercury (II) cation to T:T mismatch base pair in heteroduplex could be a convenient strategy for heteroduplex analysis and may eventually lead to progress in single nucleotide polymorphism genotyping.

Base Pair Mismatch↗
PubMed2003

Promotion of triplex formation by morpholino modification: thermodynamic and kinetic studies.

We examined the effect of morpholino (MOR) backbone modification of triplex-forming oligonucleotide (TFO) on the pyrimidine motif triplex formation at neutral pH, a condition where pyrimidine motif triplexes are unstable. The binding constant of the pyrimidine motif triplex formation at pH 6.8 with MOR-modified TFO was approximately 60 times larger than that observed with unmodified TFO. Kinetic data demonstrated that the observed increase in the binding constant at neutral pH by the MOR backbone modification resulted mainly from the considerable increase in the association rate constant. The present results certainly support the idea that the MOR backbone modification of TFO could be a key modification and may eventually lead to progress in therapeutic applications of the antigene strategy in vivo.

Base Sequence↗
PubMed2003

Thermodynamic analyses of purine motif triplex DNA formation.

We analyzed the thermodynamics of purine motif triplex formations involving single mismatches of four different base triplets (A x A:T, T x A:T, A x T:A and T x T:A) by isothermal titration calorimetry. The T x A:T and A x A:T base triplets are the most stable and the A x T:A base triplet is the least stable among the four base triplets. The magnitude of the Gibbs free energy change varies within 1.0 kcal mol(-1) for the single mismatches of the base triplet. On the other hand, the magnitude of the enthalpy change exhibits very large sequence dependence with up to 14 kcal mol(-1) variation per the single mismatches among the four base triplets, which is 14 times larger than the sequence-dependent variation of the magnitude of the Gibbs free energy change. Thus, the single mismatches in the purine motif triplex do not have strong effects on the magnitude of the Gibbs free energy change, but the magnitude of the enthalpy change exhibits much larger sequence-dependent variations. The present results support the previously proposed nucleation-elongation mechanism of the triplex formation and offer some hints to design novel triplex-forming oligonucleotides with enhanced sequence specificity.

DNA↗
PubMed2003

Rational combination of strategies to achieve synergistic stabilization of triplex.

The combination of two stabilizing strategies to increase stability of nucleic acid assembly is not always resulted in synergistic effect. In the present study, to explore the rational way to select the combination for synergistically stabilizing strategies, we examined the kinetic effects of different triplex-stabilizing strategies, poly(L-lysine)-graft-dextran (PLL-g-Dex) copolymer and N3'-->P5' phosphoramidate (PN) backbone modification of triplex-forming oligonucleotide (TFO). The former increased the association rate constant, whereas the latter decreased the dissociation rate constant in triplex equilibrium. Each strategy increased the binding constant of the triplex formation by nearly two orders of magnitude. The combination of both stabilizing strategies, which was the triplex formation with PN TFO in the presence of the copolymer, increased the binding constant by nearly four orders of magnitude, implying successful synergy of their activities. The kinetically orchestrated effects in which the copolymer and the PN modification contribute to distinct ingredients in triplex equilibrium achieved the observed synergistic stabilization. We conclude that kinetic analyses of stabilizing effects enable us to select a rational combination of stabilizing strategies.

Base Sequence↗