記述言語：英語   掲載種別：研究論文（学術雑誌）  

Fully modified 4'-thioDNA, an oligonucleotide only comprising 2'-deoxy-4'-thionucleosides, exhibited resistance to an endonuclease, in addition to preferable hybridization with RNA. Therefore, 4'-thioDNA is promising for application as a functional oligonucleotide. Fully modified 4'-thioDNA was found to behave like an RNA molecule, but no details of its structure beyond the results of circular dichroism analysis are available. Here, we have determined the structure of fully modified 4'-thioDNA with the sequence of d(CGCGAATTCGCG) by NMR. Most sugars take on the C3'-endo conformation. The major groove is narrow and deep, while the minor groove is wide and shallow. Thus, fully modified 4'-thioDNA takes on the A-form characteristic of RNA, both locally and globally. The only structure reported for 4'-thioDNA showed that partially modified 4'-thioDNA that contained some 2'-deoxy-4'-thionucleosides took on the B-form in the crystalline form. We have determined the structure of 4'-thioDNA in solution for the first time, and demonstrated unexpected differences between the two structures. The origin of the formation of the A-form is discussed. The remarkable biochemical properties reported for fully modified 4'-thioDNA, including nuclease-resistance, are rationalized in the light of the elucidated structure.

DOI： 10.1093/nar/gkn011

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Musashi protein is supposed to be involved in the regulation of differentiation of neural stem cells. Musashi binds to 3' untranslated region of target mRNA and represses the translation of mRNA. Musashi has two tandem RNA-binding domains (RBDs), RBD1 and RBD2. Both RBDs cooperatively bind to the target mRNA. Here, we determined the structure of RBD1-RBD2 in complex with target RNA. First, the structures of two RBDs in the complex were determined on the basis of short-range distance restrains derived from NOEs. However, the relative position of two RBDs was not determined due to the lack of long-range distance restraints across two RBDs. In order to overcome the situation, we introduced the paramagnetic center into Musashi by attaching MTSL carrying the NO radical. The long-range distance restraints (ca. 20-40 A) between two RBDs were derived from paramagnetic relaxation enhancement (PRE) caused by the paramagnetic center. The relative position of two RBDs was successfully determined on the basis of these long-range distance restraints. The change in the relative position of two RBDs on binding to the target RNA was also detected by PRE. The determined structure of RBD1-RBD2 in the complex has suggested how Musashi recognizes its target mRNA.

DOI： 10.1093/nass/nrn098

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

4'-thio DNA consisting of 2'-deoxy-4'-thionucleosides exhibits resistance to both endonuclease and 3'-exonuclease cleavages. Interestingly, we found that 4'-thioDNA duplex behaved like RNA molecules in hybridization properties and structural aspects. Here, we have determined the structure of 4'-thioDNA duplex in solution by NMR. Most residues take on C3'-endosugar puckering, which is characteristic to A-form. The major groove of 4'-thioDNA duplex is narrow and deep, while the minor groove is wide and shallow. These features are also characteristic to A-form. Thus, although DNA duplex usually takes on B-form in solution, 4'-thioDNA takes on A-form, like RNA molecules. A-form-like groove features of 4'-thioDNA can account for the fact that 4'-thioDNA duplex interacts with an RNA major groove binder, but not with DNA groove binder. Interaction of 4'-thioDNA with RNase V1 and resistance to endonuclease may also be accounted for in the same context.

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Interactions with DNA and RNA of three different proteins involved in the regulation of (1) transcription, (2) translation, and (3) telomere elongation were examined by NMR. In the first case, the combination of structural determination, dynamical analysis on the basis of relaxation data and identification of interactive surface for wild and phosphorylation-mimicking mutant proteins has given the insight on the increase of DNA-binding affinity through phosphorylation of the protein. In the second case, the arrangement of two tandem domains interacting with RNA has been determined with residual dipolar couplings and paramagnetic relaxation enhancement, which has given the idea on how the two tandem domains recognize the target RNA. In the third case, simultaneous binding of the other two tandem domains to both DNA and RNA has been analyzed with chemical shift perturbation analysis. The result has suggested that the protein composed of two tandem domains can recruit telomerase to telomere DNA.

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

A water-soluble cationic porphyrin, 5,10,15,20-tetrakis(N-methylpyridinium-4-yl)-21H,23H-porphyrin (TmPyP4), has been studied extensively because of its unique physicochemical properties that lead to interactions with nucleic acids, as well as its therapeutic application. Formation of a complex between TmPyP4 and parallel G-quadruplex DNA formed from a single repeat sequence of the human telomere, d(TTAGGG), has been characterized in an effort to elucidate the mode of molecular recognition between TmPyP4 and the DNA. The study demonstrated that TmPyP4 intercalates into the A3pG4 step of [d(TTAGGG)]4 with an association constant of 6.2 x 10(6) M(-1) and a stoichiometric ratio of 1:1. The binding of TmPyP4 to the A3pG4 step of [d(TTAGGG)]4 was found to be stabilized by the pi-pi stacking interaction of the porphyrin ring of TmPyP4 with the G4 quartet as well as the A3 bases of the G-quadruplex DNA. These findings provide novel insights for the design of porphyrin derivatives that bind to DNA with high affinity and specificity.

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

We reported that distamycin A can bind to DNA containing the (6-4) photoproduct, one of the major UV lesions in DNA, by means of CD spectroscopy. Here, we have analyzed the structure of DNA containing the (6-4) photoproduct in complex with distamycin A by NMR. Broadening of NMR resonances and large chemical shift perturbation have been observed for residues located close to the (6-4) photoproduct in DNA, indicating the binding to this region. Intermolecular NOEs between DNA and distamycin A have been identified for some of these residues, suggesting the formation of the relatively tight complex. On the basis of these NOEs, structure determination of the complex is now in progress.

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Parallel G-quadruplexes formed from oligonucleotide sequences, d(TTAGn), where n = 3-5, have been shown to form a dimer through end-to-end stacking of 3'-terminal G-tetrads. The monomers and dimers of the G-quadruplexes are in dynamic equilibrium with an exchange rate of approximately 1 s-1. A thermodynamic study demonstrated that the dimerization of the G-quadruplexes is largely enthalpic in origin.

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Characterization of the interaction between DNA and small organic compounds is of considerable importance for gaining insights into the mechanism underlying molecular recognition, which could be highly relevant to drug design. In the present study, the interaction of a water-soluble cationic porphyrin, 5,10,15,20-tetrakis(N-methylpyridinium-4-yl)-21H,23H-porphyrin, with a self-complementary duplex DNA, d(GCTTAAGC)2, has been investigated by means of absorption, circular dichroism, and NMR spectroscopies. The optical studies indicated that TMPyP binds to the TTAA region of d(GCTTAAGC)2 with a binding constant of 2.5 x 10(6) M(-1) and a stoichiometric ratio of 1:1. The observation of intermolecular nuclear Overhauser effect connectivities demonstrated that TMPyP binds in the major groove of d(GCTTAAGC)2. A model for the binding of TMPyP in the major groove of the AT-rich region of d(GCTTAAGC)2 is proposed.

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DNA fragments of d(TTAG(n)) (n = 3-5) form all-parallel G-quadruplexed structure in the presence of K+. We found that G-quadruplexed d(TTAG(n)) forms dimer through end-to-end stacking of the 3'-terminal G-tetrads. In this study, we report the structure of the dimer, and the dynamics and thermodynamics of the dimerization.

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

We analyzed the coordination structure of G-quadruplexed DNA-heme complex which exhibits remarkably similar spectroscopic characters to hemoprotein such as myoglobin. We found that some exogenous ligands can be accommodated at the 6th coordination site of heme in the G-quadruplexed DNA-heme complex with a DNA base coordinated to the 5th site. The G-quadruplexed DNA-hemin complex is expected to play a role as a novel DNAzyme. These findings provide novel insights into molecular design for creating artificial heme enzymes using spontaneous assembly between G-quadruplexed DNA and heme.

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

A water-soluble cationic porphyrin, 5,10,15,20-Tetra(N-methylpyridinium-4-yl)-21H,23H-porphyrin has been shown to intercalate selectively into the A3-G4 gap of C-quadruplexed DNA d(TTAGGG)4.

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Haemin, iron(III)-protoporphyrin IX complex, and parallel-quadruplexed d(TTAGGG) have been shown to form a stable coordination complex which exhibits spectroscopic properties remarkably similar to those of haemoproteins.

PubMed

researchmap

記述言語：英語   掲載種別：研究論文（学術雑誌）  

5,10,15,20-Tetra(N-methyl-pyridinium-4-yl)-21H,23H-porphyrin has shown to bind to the major groove of AT-rich DNA predominantly through electrostatic interaction between positively charged N-methyl pyridinium moieties of the porphyrin and negatively charged phosphodiester backbone. Solution structure of the complex between the porphyrin and a double-stranded DNA fragment has been inferred from the measurements of the mixing time-dependent intermolecular nuclear Overhauser effects (NOEs).

PubMed

researchmap