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

Ryo Iwaoka, Takashi Nagata, Kengo Tsuda, Takao Imai, Hideyuki Okano, Naohiro Kobayashi, Masato Katahira

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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.

Original languageEnglish
JournalMolecules (Basel, Switzerland)
Volume22
Issue number7
DOIs
Publication statusPublished - 2017 Jul 19
Externally publishedYes

Fingerprint

RNA
Messenger RNA
Structural Models
Hydrogen Bonding
Carcinogenesis
Stem Cells
Maintenance
Observation
Chemical shift
Stem cells
Conformations
stem cells
Hydrogen bonds
helices
maintenance
chemical equilibrium
perturbation
RNA-Binding Motifs
hydrogen
interactions

Keywords

  • Msi1
  • protein-RNA complex
  • RNA-binding protein
  • solution structure determination

ASJC Scopus subject areas

  • Analytical Chemistry
  • Chemistry (miscellaneous)
  • Molecular Medicine
  • Pharmaceutical Science
  • Drug Discovery
  • Physical and Theoretical Chemistry
  • Organic Chemistry

Cite this

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. / Iwaoka, Ryo; Nagata, Takashi; Tsuda, Kengo; Imai, Takao; Okano, Hideyuki; Kobayashi, Naohiro; Katahira, Masato.

In: Molecules (Basel, Switzerland), Vol. 22, No. 7, 19.07.2017.

Research output: Contribution to journalArticle

Iwaoka, Ryo ; Nagata, Takashi ; Tsuda, Kengo ; Imai, Takao ; Okano, Hideyuki ; Kobayashi, Naohiro ; Katahira, Masato. / 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. In: Molecules (Basel, Switzerland). 2017 ; Vol. 22, No. 7.
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