論文 - 三好 大輔
-
Investigation of the physicochemical and functional properties of poly(2-methacryloyloxyethyl phosphorylcholine)-conjugated aptamers. 国際誌
Seojung Cho, Jumpei Morimoto, Yutaro Saito, Yukiko Nagai, Asuka Sakata, Keitaro Yoshimoto, Mitsuki Tsuruta, Daisuke Miyoshi, Shinsuke Sando
Biomaterials science 14 ( 1 ) 232 - 239 2026年1月
Polymer conjugation is a common strategy to improve the pharmacokinetics of aptamers, yet its effects on aptamer properties are incompletely understood. Poly(ethylene glycol) (PEG) is the most widely used polymer for this purpose, but concerns about anti-PEG immune responses have prompted interest in alternative polymers. We previously reported that conjugation with the zwitterionic polymer poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) significantly prolongs the circulation time of a DNA aptamer while avoiding anti-PEG antibody recognition. In this study, we evaluated the physicochemical and functional consequences of PMPC conjugation of aptamers. Biophysical analyses suggested that the secondary structure and target-binding affinity of the aptamer were preserved, while functional consequences upon PMPC conjugation varied with the targets. The activity of a membrane receptor-targeting aptamer partially decreased, likely due to spatial constraints around the cell membrane, while RB005, targeting soluble activated coagulation factor IX, retained its full activity. In addition, PMPC conjugation significantly prolonged the in vivo plasma retention of RB005. By elucidating the effects of PMPC on aptamer properties and introducing another example that further supports the general applicability of PMPC conjugation in enhancing aptamer pharmacokinetics, these findings support PMPC as a promising alternative to PEG.
DOI： 10.1039/d5bm01078k
-
Phase Separation of RX Repeat Peptides with Nucleic Acids. 国際誌
Sumit Shil, Mitsuki Tsuruta, Ryosuke Suzuki, Yoshiki Hashimoto, Takeru Torii, Shinya Taniguchi, Tomohiro Umetani, Keiko Kawauchi, Daisuke Miyoshi
Chemistry, an Asian journal 20 ( 23 ) e00805 2025年12月
Biomolecular liquid-liquid phase separation (LLPS) plays a crucial role in organizing membraneless cellular compartments, which regulate a wide variety of cellular processes. A key molecular mechanism underlying LLPS of nucleic acids involves G-quadruplex (G4) structures of DNA and RNA interacting with intrinsically disordered proteins, particularly arginine and glycine (RGG/RG) rich proteins. The role of arginine residues in LLPS has been studied extensively, whereas few studies have focused on the role of the another frequently occurring residues, glycine. Here, we systematically investigated the contribution of G residues by substituting them with alanine (A), proline (P), valine (V), and tyrosine (Y) residues, generating a series of RX repeat peptides. Turbidity and microscopy assays with DNA oligonucleotides forming G4, duplex, as well as random coil, showed that RP and RA-peptides enhanced LLPS with G4 DNA, by comparing RG-peptide. In contrast, RY promoted liquid-solid phase separation (LSPS) with the G4 DNA, although it underwent LLPS with the random coil and duplex DNAs. In addition, RV-peptide formed aggregates even in the absence of any DNA. These results demonstrate that side-chain size, hydrophobicity, and aromaticity are critical factors for the LLPS and LSPS capability and selectivity with DNA forming various secondary structures. This study provides mechanistic insights into protein-nucleic acid LLPS and LSPS and guides the rational design peptides to undergo LLPS but not LSPS with nucleic acids.
-
Liquid-liquid phase separation induced by i-motif DNA under molecular crowding conditions 招待あり 査読あり
Ryosuke Suzuki, Mitsuki Tsuruta, Sumit Shil, Kosei Morohashi, Keiko Kawauchi, Daisuke Miyoshia
Polymer J. 57 931 - 940 2025年4月
担当区分：責任著者
-
Three- and four-stranded nucleic acid structures and their ligands 招待あり 査読あり
Yoshiki Hashimoto, Sumit Shil, Mitsuki Tsuruta, Keiko Kawauchi, and Daisuke Miyoshi
RSC Chemical Biology 6 466 - 491 2025年3月
担当区分：責任著者
DOI： 10.1039/D4CB00287C
-
ケミカルバイオロジーの冒険（１２） 核酸を標的とした低分子・中分子化合物 招待あり
堂野主税、勝田陽介、三好大輔
現代化学 648 32 - 39 2025年2月
担当区分：最終著者
-
Roles of Loop Region in Folding Kinetics and Transcription Inhibition of DNA G-Quadruplexes 査読あり
Minori Nakata, Naoki Kosaka, Keiko Kawauchi, Daisuke Miyoshi
Biochemistry 64 609 - 619 2025年1月
担当区分：責任著者
-
Heterogeneity of Molecular Crowding and Liquid-Liquid Phase Separation. 国際誌
Mitsuki Tsuruta, Sumit Shil, Keiko Kawauchi, Daisuke Miyoshi
Sub-cellular biochemistry 109 327 - 345 2025年
The inside of a living cell is highly crowded with extremely diverse biomacromolecules, small metabolites and osmolytes. The molecular conditions in cells change dynamically and rapidly depending on the cell cycle and state, organelle, and compartment. Much remains unknown regarding how biomolecular interactions and reactions can proceed in a spatiotemporally specific manner in such crowded, heterogeneous, and dynamic molecular environments. Selective condensation/droplet formation of biomolecules via liquid-liquid phase separation may be critical for interactions and reactions inside cells. In this chapter, we briefly describe the heterogeneity of molecular environments inside cells and the biological roles of liquid-liquid phase separation that allows biomolecular interactions and reactions in such heterogenous molecular environments. Finally, we discuss the mutual relationship between molecular crowding and liquid-liquid phase separation.
-
NRAS DNA G-quadruplex-targeting molecules for sequence-selective enzyme inhibition 招待あり 査読あり
Yoshiki Hashimoto, Hiroki Kubo, Keiko Kawauchi and Daisuke Miyoshi
Chem. Commun. 60 13179 - 13182 2024年10月
担当区分：責任著者
DOI： 10.1039/D4CC03753G
-
The role of cytosine methylation in regulating the topology and liquid-liquid phase separation of DNA G-quadruplexes 査読あり
Mitsuki Tsuruta, Sumit Shil, Shinya Taniguchi, Keiko Kawauchi and Daisuke Miyoshi
Chem. Sci. 16 4213 - 4225 2024年10月
担当区分：責任著者
DOI： 10.1039/D4SC06959E
-
Factors Affecting Liquid-Liquid Phase Separation of RGG Peptides with DNA G-Quadruplex 招待あり 査読あり
Sumit Shil, Mitsuki Tsuruta, Keiko Kawauchi, Daisuke Miyoshi
ChemMedChem 2024年9月
-
Bioinformatic Analysis of Actin-Binding Proteins in the Nucleolus During Heat Shock 招待あり 査読あり
Shinya Taniguchi, Takeru Torii ,Toshiyuki Goto, Kohei Takeuchi, Rine Katsumi, Mako Sumida, Sunmin Lee, Wataru Sugimoto, Masaya Gessho, Katsuhiko Itoh, Hiroaki Hirata, Junji Kawakami, Daisuke Miyoshi and Keiko Kawauchi
Genes 15 ( 12 ) 1580 2024年9月
-
Quantitative Effects of the Loop Region on Topology, Thermodynamics, and Cation Binding of DNA G-quadruplexes 査読あり
Minori Nakata, Naoki Kosaka, Keiko Kawauchi, Daisuke Miyoshi
ACS Omega 9 35028 - 35036 2024年7月
担当区分：最終著者, 責任著者
-
細胞内環境で安定化する核酸構造を標的にした低分子薬の開発 招待あり
橋本 佳樹, 川内 敬子, 三好 大輔
MEDCHEM NEWS (日本薬学会） 34 ( 1 ) 36 - 42 2024年4月
担当区分：責任著者
-
RNA 高次構造を標的とした光線力学療法の展望 招待あり
取井猛流・木下菜月・橋本佳樹・杉本渉・川内敬子・三 好大輔
38 ( 13 ) 1154 - 1156 2023年11月
担当区分：最終著者, 責任著者
-
Loss of p53 function promotes DNA damage-induced formation of nuclear actin filaments 査読あり 国際誌
Takeru Torii, Wataru Sugimoto, Katsuhiko Itoh, Natsuki Kinoshita, Masaya Gessho, Toshiyuki Goto, Ikuno Uehara, Wataru Nakajima, Yemima Budirahardja, Daisuke Miyoshi, Takahito Nishikata, Nobuyuki Tanaka, Hiroaki Hirata, Keiko Kawauchi
Cell Death & Disease 14 ( 11 ) 766766 - 766 2023年8月
Tumor suppressor p53 plays a central role in response to DNA damage. DNA-damaging agents modulate nuclear actin dynamics, influencing cell behaviors; however, whether p53 affects the formation of nuclear actin filaments remains unclear. In this study, we found that p53 depletion promoted the formation of nuclear actin filaments in response to DNA-damaging agents, such as doxorubicin (DOXO) and etoposide (VP16). Even though the genetic probes used for the detection of nuclear actin filaments exerted a promotive effect on actin polymerization, the detected formation of nuclear actin filaments was highly dependent on both p53 depletion and DNA damage. Whilst active p53 is known to promote caspase-1 expression, the overexpression of caspase-1 reduced DNA damage-induced formation of nuclear actin filaments in p53-depleted cells. In contrast, co-treatment with DOXO and the pan-caspase inhibitor Q-VD-OPh or the caspase-1 inhibitor Z-YVAD-FMK induced the formation of nuclear actin filament formation even in cells bearing wild-type p53. These results suggest that the p53-caspase-1 axis suppresses DNA damage-induced formation of nuclear actin filaments. In addition, we found that the expression of nLifeact-GFP, the filamentous-actin-binding peptide Lifeact fused with the nuclear localization signal (NLS) and GFP, modulated the structure of nuclear actin filaments to be phalloidin-stainable in p53-depleted cells treated with the DNA-damaging agent, altering the chromatin structure and reducing the transcriptional activity. The level of phosphorylated H2AX (γH2AX), a marker of DNA damage, in these cells also reduced upon nLifeact-GFP expression, whilst details of the functional relationship between the formation of nLifeact-GFP-decorated nuclear actin filaments and DNA repair remained to be elucidated. Considering that the loss of p53 is associated with cancer progression, the results of this study raise a possibility that the artificial reinforcement of nuclear actin filaments by nLifeact-GFP may enhance the cytotoxic effect of DNA-damaging agents in aggressive cancer cells through a reduction in gene transcription.
-
The iron chelator deferriferrichrysin induces paraptosis via extracellular signal-related kinase activation in cancer cells 招待あり 査読あり
Natsuki Kinoshita, Masaya Gessho, Takeru Torii, Yukako Ashida, Minori Akamatsu, Alvin Kunyao Guo, Sunmin Lee, Tatsuya Katsuno 3, Wataru Nakajima Yemima Budirahardja, Daisuke Miyoshi, Takehiko Todokoro, Hiroki Ishida, Takahito Nishikata, Keiko Kawauchi
Genes Cells 28 ( 9 ) 653 - 662 2023年6月
DOI： 10.1111/gtc.13053
-
Quantification of the concentration in a droplet formed by liquid–liquid phase separation of G-quadruplex-forming RNA 査読あり
Kohei Yokosawa, Mitsuki Tsuruta, Shinji Kajimoto, Naoki Sugimoto, Daisuke Miyoshi, Takakazu Nakabayashi
Chem. Phys. Lett. 826 140634 - 140634 2023年6月
-
Biomolecular Liquid–Liquid Phase Separation for Biotechnology 招待あり 査読あり
Sumit Shil, Mitsuki Tsuruta, Keiko Kawauchi and Daisuke Miyoshi
BioTech 12 ( 2 ) 26 2023年3月
担当区分：責任著者
-
グアニン四重らせん構造による転移因子LINE-1の発現制御 招待あり
月生 雅也, 李 先民, Yemima Suryani Budirahardja, 鶴田 充生, 橋本 佳樹, 高宮 渚, 木下 菜月, 建石 寿枝, 杉本 直己, 三好 大輔, 川内 敬子
日本女性科学者の会学術誌 23 48 - 48 2023年3月
出版者・発行元：(一社)日本女性科学者の会
-
Simple and fast screening for structure-selective G-quadruplex ligands 査読あり 国際誌
Yoshiki Hashimoto, Yoshiki Imagawa, Kaho Nagano, Ryuichi Maeda, Naho Nagahama, Takeru Torii, Natsuki Kinoshita, Nagisa Takamiya, Keiko Kawauchi, Hisae Tatesishi-Karimata, Naoki Sugimoto and Daisuke Miyoshi
Chem. Commun. 59 ( 33 ) 4891 - 4894 2023年2月
担当区分：責任著者
Structural selectivity of G-quadruplex ligands is essential for cellular applications since there is an excess of nucleic acids forming duplex structures compared to G-quadruplex structures in living cells. In this study, we developed new structure-selective G-quadruplex ligands utilizing a simple and fast screening system. The affinity, selectivity, enzymatic inhibitory activity and cytotoxicity of the structure-selective G-quadruplex ligands were demonstrated along with a structural selectivity-cytotoxicity relationship of G-quadruplex ligands.
DOI： 10.1039/D3CC00556A