Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA
Musashi-1 (Msi1) controls the maintenance of stem cells and tumorigenesis through binding to its target mRNAs and subsequent translational regulation. Msi1 has two RNA-binding domains (RBDs), RBD1 and RBD2, which recognize r(GUAG) and r(UAG), respectively. These minimal recognition sequences are con...
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MDPI AG
2017-07-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/22/7/1207 |
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| author | Ryo Iwaoka Takashi Nagata Kengo Tsuda Takao Imai Hideyuki Okano Naohiro Kobayashi Masato Katahira |
| author_facet | Ryo Iwaoka Takashi Nagata Kengo Tsuda Takao Imai Hideyuki Okano Naohiro Kobayashi Masato Katahira |
| author_sort | Ryo Iwaoka |
| collection | DOAJ |
| description | Musashi-1 (Msi1) controls the maintenance of stem cells and tumorigenesis through binding to its target mRNAs and subsequent translational regulation. Msi1 has two RNA-binding domains (RBDs), RBD1 and RBD2, which recognize r(GUAG) and r(UAG), respectively. These minimal recognition sequences are connected by variable linkers in the Msi1 target mRNAs, however, the molecular mechanism by which Msi1 recognizes its targets is not yet understood. We previously determined the solution structure of the Msi1 RBD1:r(GUAGU) complex. Here, we determined the first structure of the RBD2:r(GUAGU) complex. The structure revealed that the central trinucleotide, r(UAG), is specifically recognized by the intermolecular hydrogen-bonding and aromatic stacking interactions. Importantly, the C-terminal region, which is disordered in the free form, took a certain conformation, resembling a helix. The observation of chemical shift perturbation and intermolecular NOEs, together with increases in the heteronuclear steady-state {1H}-15N NOE values on complex formation, indicated the involvement of the C-terminal region in RNA binding. On the basis of the two complex structures, we built a structural model of consecutive RBDs with r(UAGGUAG) containing both minimal recognition sequences, which resulted in no steric hindrance. The model suggests recognition of variable lengths (n) of the linker up to n = 50 may be possible. |
| first_indexed | 2024-12-10T08:51:07Z |
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| id | doaj.art-e1889a951fe14e66ba680d2af492d6e1 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-10T08:51:07Z |
| publishDate | 2017-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-e1889a951fe14e66ba680d2af492d6e12022-12-22T01:55:36ZengMDPI AGMolecules1420-30492017-07-01227120710.3390/molecules22071207molecules22071207Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNARyo Iwaoka0Takashi Nagata1Kengo Tsuda2Takao Imai3Hideyuki Okano4Naohiro Kobayashi5Masato Katahira6Institute of Advanced Energy, Kyoto University, Gokasho, Uji, Kyoto 611-0011, JapanInstitute of Advanced Energy, Kyoto University, Gokasho, Uji, Kyoto 611-0011, JapanRIKEN Center for Life Science Technologies, 1-7-22 Suehirocho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, JapanDepartment of Physiology, Keio University School of Medicine, Shinanomachi, Shinjuku-ku, Tokyo 160-8582, JapanDepartment of Physiology, Keio University School of Medicine, Shinanomachi, Shinjuku-ku, Tokyo 160-8582, JapanInstitute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, JapanInstitute of Advanced Energy, Kyoto University, Gokasho, Uji, Kyoto 611-0011, JapanMusashi-1 (Msi1) controls the maintenance of stem cells and tumorigenesis through binding to its target mRNAs and subsequent translational regulation. Msi1 has two RNA-binding domains (RBDs), RBD1 and RBD2, which recognize r(GUAG) and r(UAG), respectively. These minimal recognition sequences are connected by variable linkers in the Msi1 target mRNAs, however, the molecular mechanism by which Msi1 recognizes its targets is not yet understood. We previously determined the solution structure of the Msi1 RBD1:r(GUAGU) complex. Here, we determined the first structure of the RBD2:r(GUAGU) complex. The structure revealed that the central trinucleotide, r(UAG), is specifically recognized by the intermolecular hydrogen-bonding and aromatic stacking interactions. Importantly, the C-terminal region, which is disordered in the free form, took a certain conformation, resembling a helix. The observation of chemical shift perturbation and intermolecular NOEs, together with increases in the heteronuclear steady-state {1H}-15N NOE values on complex formation, indicated the involvement of the C-terminal region in RNA binding. On the basis of the two complex structures, we built a structural model of consecutive RBDs with r(UAGGUAG) containing both minimal recognition sequences, which resulted in no steric hindrance. The model suggests recognition of variable lengths (n) of the linker up to n = 50 may be possible.https://www.mdpi.com/1420-3049/22/7/1207RNA-binding proteinMsi1solution structure determinationprotein-RNA complex |
| spellingShingle | Ryo Iwaoka Takashi Nagata Kengo Tsuda Takao Imai Hideyuki Okano Naohiro Kobayashi Masato Katahira Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA Molecules RNA-binding protein Msi1 solution structure determination protein-RNA complex |
| title | Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA |
| title_full | Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA |
| title_fullStr | Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA |
| title_full_unstemmed | Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA |
| title_short | Structural Insight into the Recognition of r(UAG) by Musashi-1 RBD2, and Construction of a Model of Musashi-1 RBD1-2 Bound to the Minimum Target RNA |
| title_sort | structural insight into the recognition of r uag by musashi 1 rbd2 and construction of a model of musashi 1 rbd1 2 bound to the minimum target rna |
| topic | RNA-binding protein Msi1 solution structure determination protein-RNA complex |
| url | https://www.mdpi.com/1420-3049/22/7/1207 |
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