The neural RNA-binding protein musashi1 translationally regulates mammalian numb gene expression by interacting with its mRNA

Takao Imai, Akinori Tokunaga, Tetsu Yoshida, Mitsuhiro Hashimoto, Katsuhiko Mikoshiba, Gerry Weinmaster, Masato Nakafuku, Hideyuki Okano

Research output: Contribution to journalArticle

310 Citations (Scopus)

Abstract

Musashi1 (Msi1) is an RNA-binding protein that is highly expressed in neural progenitor cells, including neural stem cells. In this study, the RNA-binding sequences for Msi1 were determined by in vitro selection using a pool of degenerate 50-mer sequences. All of the selected RNA species contained repeats of (G/A)UnAGU (n = 1 to 3) sequences which were essential for Msi1 binding. These consensus elements were identified in some neural mRNAs. One of these, mammalian numb (m-numb), which encodes a membrane-associated antagonist of Notch signaling, is a likely target of Msi1. Msi1 protein binds in vitro-transcribed m-numb RNA in its 3′-untranslated region (UTR) and binds endogenous m-numb mRNA in vivo, as shown by affinity precipitation followed by reverse transcription-PCR. Furthermore, adenovirus-induced Msi1 expression resulted in the down-regulation of endogenous m-Numb protein expression. Reporter assays using a chimeric mRNA that combined luciferase and the 3′-UTR of m-numb demonstrated that Msi1 decreased the reporter activity without altering the reporter mRNA level. Thus, our results suggested that Msi1 could regulate the expression of its target gene at the translational level. Furthermore, we found that Notch signaling activity was increased by Msi1 expression in connection with the posttranscriptional down-regulation of the m-numb gene.

Original languageEnglish
Pages (from-to)3888-3900
Number of pages13
JournalMolecular and Cellular Biology
Volume21
Issue number12
DOIs
Publication statusPublished - 2001 Jun

Fingerprint

RNA-Binding Proteins
Gene Expression
Messenger RNA
3' Untranslated Regions
Down-Regulation
RNA
Neural Stem Cells
Luciferases
Adenoviridae
Genes
Reverse Transcription
Proteins
Stem Cells
Polymerase Chain Reaction
Membranes
In Vitro Techniques

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

The neural RNA-binding protein musashi1 translationally regulates mammalian numb gene expression by interacting with its mRNA. / Imai, Takao; Tokunaga, Akinori; Yoshida, Tetsu; Hashimoto, Mitsuhiro; Mikoshiba, Katsuhiko; Weinmaster, Gerry; Nakafuku, Masato; Okano, Hideyuki.

In: Molecular and Cellular Biology, Vol. 21, No. 12, 06.2001, p. 3888-3900.

Research output: Contribution to journalArticle

Imai, Takao ; Tokunaga, Akinori ; Yoshida, Tetsu ; Hashimoto, Mitsuhiro ; Mikoshiba, Katsuhiko ; Weinmaster, Gerry ; Nakafuku, Masato ; Okano, Hideyuki. / The neural RNA-binding protein musashi1 translationally regulates mammalian numb gene expression by interacting with its mRNA. In: Molecular and Cellular Biology. 2001 ; Vol. 21, No. 12. pp. 3888-3900.
@article{17ec5764c3764e9191b7514278f41def,
title = "The neural RNA-binding protein musashi1 translationally regulates mammalian numb gene expression by interacting with its mRNA",
abstract = "Musashi1 (Msi1) is an RNA-binding protein that is highly expressed in neural progenitor cells, including neural stem cells. In this study, the RNA-binding sequences for Msi1 were determined by in vitro selection using a pool of degenerate 50-mer sequences. All of the selected RNA species contained repeats of (G/A)UnAGU (n = 1 to 3) sequences which were essential for Msi1 binding. These consensus elements were identified in some neural mRNAs. One of these, mammalian numb (m-numb), which encodes a membrane-associated antagonist of Notch signaling, is a likely target of Msi1. Msi1 protein binds in vitro-transcribed m-numb RNA in its 3′-untranslated region (UTR) and binds endogenous m-numb mRNA in vivo, as shown by affinity precipitation followed by reverse transcription-PCR. Furthermore, adenovirus-induced Msi1 expression resulted in the down-regulation of endogenous m-Numb protein expression. Reporter assays using a chimeric mRNA that combined luciferase and the 3′-UTR of m-numb demonstrated that Msi1 decreased the reporter activity without altering the reporter mRNA level. Thus, our results suggested that Msi1 could regulate the expression of its target gene at the translational level. Furthermore, we found that Notch signaling activity was increased by Msi1 expression in connection with the posttranscriptional down-regulation of the m-numb gene.",
author = "Takao Imai and Akinori Tokunaga and Tetsu Yoshida and Mitsuhiro Hashimoto and Katsuhiko Mikoshiba and Gerry Weinmaster and Masato Nakafuku and Hideyuki Okano",
year = "2001",
month = "6",
doi = "10.1128/MCB.21.12.3888-3900.2001",
language = "English",
volume = "21",
pages = "3888--3900",
journal = "Molecular and Cellular Biology",
issn = "0270-7306",
publisher = "American Society for Microbiology",
number = "12",

}

TY - JOUR

T1 - The neural RNA-binding protein musashi1 translationally regulates mammalian numb gene expression by interacting with its mRNA

AU - Imai, Takao

AU - Tokunaga, Akinori

AU - Yoshida, Tetsu

AU - Hashimoto, Mitsuhiro

AU - Mikoshiba, Katsuhiko

AU - Weinmaster, Gerry

AU - Nakafuku, Masato

AU - Okano, Hideyuki

PY - 2001/6

Y1 - 2001/6

N2 - Musashi1 (Msi1) is an RNA-binding protein that is highly expressed in neural progenitor cells, including neural stem cells. In this study, the RNA-binding sequences for Msi1 were determined by in vitro selection using a pool of degenerate 50-mer sequences. All of the selected RNA species contained repeats of (G/A)UnAGU (n = 1 to 3) sequences which were essential for Msi1 binding. These consensus elements were identified in some neural mRNAs. One of these, mammalian numb (m-numb), which encodes a membrane-associated antagonist of Notch signaling, is a likely target of Msi1. Msi1 protein binds in vitro-transcribed m-numb RNA in its 3′-untranslated region (UTR) and binds endogenous m-numb mRNA in vivo, as shown by affinity precipitation followed by reverse transcription-PCR. Furthermore, adenovirus-induced Msi1 expression resulted in the down-regulation of endogenous m-Numb protein expression. Reporter assays using a chimeric mRNA that combined luciferase and the 3′-UTR of m-numb demonstrated that Msi1 decreased the reporter activity without altering the reporter mRNA level. Thus, our results suggested that Msi1 could regulate the expression of its target gene at the translational level. Furthermore, we found that Notch signaling activity was increased by Msi1 expression in connection with the posttranscriptional down-regulation of the m-numb gene.

AB - Musashi1 (Msi1) is an RNA-binding protein that is highly expressed in neural progenitor cells, including neural stem cells. In this study, the RNA-binding sequences for Msi1 were determined by in vitro selection using a pool of degenerate 50-mer sequences. All of the selected RNA species contained repeats of (G/A)UnAGU (n = 1 to 3) sequences which were essential for Msi1 binding. These consensus elements were identified in some neural mRNAs. One of these, mammalian numb (m-numb), which encodes a membrane-associated antagonist of Notch signaling, is a likely target of Msi1. Msi1 protein binds in vitro-transcribed m-numb RNA in its 3′-untranslated region (UTR) and binds endogenous m-numb mRNA in vivo, as shown by affinity precipitation followed by reverse transcription-PCR. Furthermore, adenovirus-induced Msi1 expression resulted in the down-regulation of endogenous m-Numb protein expression. Reporter assays using a chimeric mRNA that combined luciferase and the 3′-UTR of m-numb demonstrated that Msi1 decreased the reporter activity without altering the reporter mRNA level. Thus, our results suggested that Msi1 could regulate the expression of its target gene at the translational level. Furthermore, we found that Notch signaling activity was increased by Msi1 expression in connection with the posttranscriptional down-regulation of the m-numb gene.

UR - http://www.scopus.com/inward/record.url?scp=0035016795&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035016795&partnerID=8YFLogxK

U2 - 10.1128/MCB.21.12.3888-3900.2001

DO - 10.1128/MCB.21.12.3888-3900.2001

M3 - Article

VL - 21

SP - 3888

EP - 3900

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 12

ER -