HMGA1a induces alternative splicing of the estrogen receptor-alpha gene by trapping U1 snRNP to an upstream pseudo-5' splice site

Kenji Ohe, Shinsuke Miyajima, Tomoko Tanaka, Yuriko Hamaguchi, Yoshihiro Harada, Yuta Horita, Yuki Beppu, Fumiaki Ito, Takafumi Yamasaki, Hiroki Terai, Masayoshi Mori, Yusuke Murata, Makito Tanabe, Ichiro Abe, Kenji Ashida, Kunihisa Kobayashi, Munechika Enjoji, Takashi Nomiyama, Toshihiko Yanase, Nobuhiro HaradaToshiaki Utsumi, Akila Mayeda

研究成果: Article

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Objectives: The high-mobility group A protein 1a (HMGA1a) protein is known as a transcription factor that binds to DNA, but recent studies have shown it exerts novel functions through RNA-binding. We were prompted to decipher the mechanism of HMGA1a-induced alternative splicing of the estrogen receptor alpha (ERα) that we recently reported would alter tamoxifen sensitivity in MCF-7 TAMR1 cells. Methods: Endogenous expression of full length ERα66 and its isoform ERα46 were evaluated in MCF-7 breast cancer cells by transient expression of HMGA1a and an RNA decoy (2'-O-methylated RNA of the HMGA1a RNA-binding site) that binds to HMGA1a. RNA-binding of HMGA1a was checked by RNA-EMSA. In vitro splicing assay was performed to check the direct involvement of HMGA1a in splicing regulation. RNA-EMSA assay in the presence of purified U1 snRNP was performed with psoralen UV crosslinking to check complex formation of HMGA1a-U1 snRNP at the upstream pseudo-5' splice site of exon 1. Results: HMGA1a induced exon skipping of a shortened exon 1 of ERα in in vitro splicing assays that was blocked by the HMGA1a RNA decoy and sequence-specific RNA-binding was confirmed by RNA-EMSA. RNA-EMSA combined with psoralen UV crosslinking showed that HMGA1a trapped purified U1 snRNP at the upstream pseudo-5' splice site. Conclusions: Regulation of ERα alternative splicing by an HMGA1a-trapped U1 snRNP complex at the upstream 5' splice site of exon 1 offers novel insight on 5' splice site regulation by U1 snRNP as well as a promising target in breast cancer therapy where alternative splicing of ERα is involved.

元の言語English
記事番号52
ジャーナルFrontiers in Molecular Biosciences
5
発行部数JUN
DOI
出版物ステータスPublished - 08-06-2018

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U1 Small Nuclear Ribonucleoproteins
High Mobility Group Proteins
RNA Splice Sites
Estrogen Receptor alpha
Alternative Splicing
Genes
RNA
Proteins
Exons
Ficusin
Assays
Crosslinking
Breast Neoplasms
MCF-7 Cells
Tamoxifen

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

これを引用

Ohe, Kenji ; Miyajima, Shinsuke ; Tanaka, Tomoko ; Hamaguchi, Yuriko ; Harada, Yoshihiro ; Horita, Yuta ; Beppu, Yuki ; Ito, Fumiaki ; Yamasaki, Takafumi ; Terai, Hiroki ; Mori, Masayoshi ; Murata, Yusuke ; Tanabe, Makito ; Abe, Ichiro ; Ashida, Kenji ; Kobayashi, Kunihisa ; Enjoji, Munechika ; Nomiyama, Takashi ; Yanase, Toshihiko ; Harada, Nobuhiro ; Utsumi, Toshiaki ; Mayeda, Akila. / HMGA1a induces alternative splicing of the estrogen receptor-alpha gene by trapping U1 snRNP to an upstream pseudo-5' splice site. :: Frontiers in Molecular Biosciences. 2018 ; 巻 5, 番号 JUN.
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title = "HMGA1a induces alternative splicing of the estrogen receptor-alpha gene by trapping U1 snRNP to an upstream pseudo-5' splice site",
abstract = "Objectives: The high-mobility group A protein 1a (HMGA1a) protein is known as a transcription factor that binds to DNA, but recent studies have shown it exerts novel functions through RNA-binding. We were prompted to decipher the mechanism of HMGA1a-induced alternative splicing of the estrogen receptor alpha (ERα) that we recently reported would alter tamoxifen sensitivity in MCF-7 TAMR1 cells. Methods: Endogenous expression of full length ERα66 and its isoform ERα46 were evaluated in MCF-7 breast cancer cells by transient expression of HMGA1a and an RNA decoy (2'-O-methylated RNA of the HMGA1a RNA-binding site) that binds to HMGA1a. RNA-binding of HMGA1a was checked by RNA-EMSA. In vitro splicing assay was performed to check the direct involvement of HMGA1a in splicing regulation. RNA-EMSA assay in the presence of purified U1 snRNP was performed with psoralen UV crosslinking to check complex formation of HMGA1a-U1 snRNP at the upstream pseudo-5' splice site of exon 1. Results: HMGA1a induced exon skipping of a shortened exon 1 of ERα in in vitro splicing assays that was blocked by the HMGA1a RNA decoy and sequence-specific RNA-binding was confirmed by RNA-EMSA. RNA-EMSA combined with psoralen UV crosslinking showed that HMGA1a trapped purified U1 snRNP at the upstream pseudo-5' splice site. Conclusions: Regulation of ERα alternative splicing by an HMGA1a-trapped U1 snRNP complex at the upstream 5' splice site of exon 1 offers novel insight on 5' splice site regulation by U1 snRNP as well as a promising target in breast cancer therapy where alternative splicing of ERα is involved.",
author = "Kenji Ohe and Shinsuke Miyajima and Tomoko Tanaka and Yuriko Hamaguchi and Yoshihiro Harada and Yuta Horita and Yuki Beppu and Fumiaki Ito and Takafumi Yamasaki and Hiroki Terai and Masayoshi Mori and Yusuke Murata and Makito Tanabe and Ichiro Abe and Kenji Ashida and Kunihisa Kobayashi and Munechika Enjoji and Takashi Nomiyama and Toshihiko Yanase and Nobuhiro Harada and Toshiaki Utsumi and Akila Mayeda",
year = "2018",
month = "6",
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doi = "10.3389/fmolb.2018.00052",
language = "English",
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journal = "Frontiers in Molecular Biosciences",
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Ohe, K, Miyajima, S, Tanaka, T, Hamaguchi, Y, Harada, Y, Horita, Y, Beppu, Y, Ito, F, Yamasaki, T, Terai, H, Mori, M, Murata, Y, Tanabe, M, Abe, I, Ashida, K, Kobayashi, K, Enjoji, M, Nomiyama, T, Yanase, T, Harada, N, Utsumi, T & Mayeda, A 2018, 'HMGA1a induces alternative splicing of the estrogen receptor-alpha gene by trapping U1 snRNP to an upstream pseudo-5' splice site', Frontiers in Molecular Biosciences, 巻. 5, 番号 JUN, 52. https://doi.org/10.3389/fmolb.2018.00052

HMGA1a induces alternative splicing of the estrogen receptor-alpha gene by trapping U1 snRNP to an upstream pseudo-5' splice site. / Ohe, Kenji; Miyajima, Shinsuke; Tanaka, Tomoko; Hamaguchi, Yuriko; Harada, Yoshihiro; Horita, Yuta; Beppu, Yuki; Ito, Fumiaki; Yamasaki, Takafumi; Terai, Hiroki; Mori, Masayoshi; Murata, Yusuke; Tanabe, Makito; Abe, Ichiro; Ashida, Kenji; Kobayashi, Kunihisa; Enjoji, Munechika; Nomiyama, Takashi; Yanase, Toshihiko; Harada, Nobuhiro; Utsumi, Toshiaki; Mayeda, Akila.

:: Frontiers in Molecular Biosciences, 巻 5, 番号 JUN, 52, 08.06.2018.

研究成果: Article

TY - JOUR

T1 - HMGA1a induces alternative splicing of the estrogen receptor-alpha gene by trapping U1 snRNP to an upstream pseudo-5' splice site

AU - Ohe, Kenji

AU - Miyajima, Shinsuke

AU - Tanaka, Tomoko

AU - Hamaguchi, Yuriko

AU - Harada, Yoshihiro

AU - Horita, Yuta

AU - Beppu, Yuki

AU - Ito, Fumiaki

AU - Yamasaki, Takafumi

AU - Terai, Hiroki

AU - Mori, Masayoshi

AU - Murata, Yusuke

AU - Tanabe, Makito

AU - Abe, Ichiro

AU - Ashida, Kenji

AU - Kobayashi, Kunihisa

AU - Enjoji, Munechika

AU - Nomiyama, Takashi

AU - Yanase, Toshihiko

AU - Harada, Nobuhiro

AU - Utsumi, Toshiaki

AU - Mayeda, Akila

PY - 2018/6/8

Y1 - 2018/6/8

N2 - Objectives: The high-mobility group A protein 1a (HMGA1a) protein is known as a transcription factor that binds to DNA, but recent studies have shown it exerts novel functions through RNA-binding. We were prompted to decipher the mechanism of HMGA1a-induced alternative splicing of the estrogen receptor alpha (ERα) that we recently reported would alter tamoxifen sensitivity in MCF-7 TAMR1 cells. Methods: Endogenous expression of full length ERα66 and its isoform ERα46 were evaluated in MCF-7 breast cancer cells by transient expression of HMGA1a and an RNA decoy (2'-O-methylated RNA of the HMGA1a RNA-binding site) that binds to HMGA1a. RNA-binding of HMGA1a was checked by RNA-EMSA. In vitro splicing assay was performed to check the direct involvement of HMGA1a in splicing regulation. RNA-EMSA assay in the presence of purified U1 snRNP was performed with psoralen UV crosslinking to check complex formation of HMGA1a-U1 snRNP at the upstream pseudo-5' splice site of exon 1. Results: HMGA1a induced exon skipping of a shortened exon 1 of ERα in in vitro splicing assays that was blocked by the HMGA1a RNA decoy and sequence-specific RNA-binding was confirmed by RNA-EMSA. RNA-EMSA combined with psoralen UV crosslinking showed that HMGA1a trapped purified U1 snRNP at the upstream pseudo-5' splice site. Conclusions: Regulation of ERα alternative splicing by an HMGA1a-trapped U1 snRNP complex at the upstream 5' splice site of exon 1 offers novel insight on 5' splice site regulation by U1 snRNP as well as a promising target in breast cancer therapy where alternative splicing of ERα is involved.

AB - Objectives: The high-mobility group A protein 1a (HMGA1a) protein is known as a transcription factor that binds to DNA, but recent studies have shown it exerts novel functions through RNA-binding. We were prompted to decipher the mechanism of HMGA1a-induced alternative splicing of the estrogen receptor alpha (ERα) that we recently reported would alter tamoxifen sensitivity in MCF-7 TAMR1 cells. Methods: Endogenous expression of full length ERα66 and its isoform ERα46 were evaluated in MCF-7 breast cancer cells by transient expression of HMGA1a and an RNA decoy (2'-O-methylated RNA of the HMGA1a RNA-binding site) that binds to HMGA1a. RNA-binding of HMGA1a was checked by RNA-EMSA. In vitro splicing assay was performed to check the direct involvement of HMGA1a in splicing regulation. RNA-EMSA assay in the presence of purified U1 snRNP was performed with psoralen UV crosslinking to check complex formation of HMGA1a-U1 snRNP at the upstream pseudo-5' splice site of exon 1. Results: HMGA1a induced exon skipping of a shortened exon 1 of ERα in in vitro splicing assays that was blocked by the HMGA1a RNA decoy and sequence-specific RNA-binding was confirmed by RNA-EMSA. RNA-EMSA combined with psoralen UV crosslinking showed that HMGA1a trapped purified U1 snRNP at the upstream pseudo-5' splice site. Conclusions: Regulation of ERα alternative splicing by an HMGA1a-trapped U1 snRNP complex at the upstream 5' splice site of exon 1 offers novel insight on 5' splice site regulation by U1 snRNP as well as a promising target in breast cancer therapy where alternative splicing of ERα is involved.

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U2 - 10.3389/fmolb.2018.00052

DO - 10.3389/fmolb.2018.00052

M3 - Article

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VL - 5

JO - Frontiers in Molecular Biosciences

JF - Frontiers in Molecular Biosciences

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