4F2hc stabilizes GLUT1 protein and increases glucose transport activity

Haruya Ohno, Yusuke Nakatsu, Hideyuki Sakoda, Akifumi Kushiyama, Hiraku Ono, Midori Fujishiro, Yuichiro Otani, Hirofumi Okubo, Masayasu Yoneda, Toshiaki Fukushima, Yoshihiro Tsuchiya, Hideaki Kamata, Fusanori Nishimura, Hiroki Kurihara, Hideki Katagiri, Yoshitomo Oka, Tomoichiro Asano

研究成果: ジャーナルへの寄稿記事

19 引用 (Scopus)

抄録

Glucose transporter 1 (GLUT1) is widely distributed throughout various tissues and contributes to insulin- independent basal glucose uptake. Using a split-ubiquitin membrane yeast two-hybrid system, we newly identified 4F2 heavy chain (4F2hc) as a membrane protein interacting with GLUT1. Though 4F2hc reportedly forms heterodimeric complexes between amino acid transporters, such as LAT1 and LAT2, and regulates amino acid uptake, we investigated the effects of 4F2hc on GLUT1 expression and the associated glucose uptake. First, FLAG-tagged 4F2hc and hemagglutinin-tagged GLUT1 were overexpressed in human embryonic kidney 293 cells and their association was confirmed by coimmunoprecipitation. The green fluorescent protein-tagged 4F2hc and DsRed-tagged GLUT1 showed significant, but incomplete, colocalization at the plasma membrane. In addition, an endogenous association between GLUT1 and 4F2hc was demonstrated using mouse brain tissue and HeLa cells. Interestingly, overexpression of 4F2hc increased the amount of GLUT1 protein in HeLa and HepG2 cells with increased glucose uptake. In contrast, small interfering RNA (siRNA)-mediated 4F2hc gene suppression markedly reduced GLUT1 protein in both cell types, with reduced glucose uptake. While GLUT1 mRNA levels were not affected by overexpression or gene silencing of 4F2hc, GLUT1 degradation after the addition of cycloheximide was significantly suppressed by 4F2hc overexpression and increased by 4F2hc siRNA treatment. Taken together, these observations indicate that 4F2hc is likely to be involved in GLUT1 stabilization and to contribute to the regulation of not only amino acid but also glucose metabolism.

元の言語英語
ページ(範囲)C1047-C1054
ジャーナルAmerican Journal of Physiology - Cell Physiology
300
発行部数5
DOI
出版物ステータス出版済み - 5 1 2011

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CD98 Heavy Chain Antigens
Facilitative Glucose Transport Proteins
Glucose
HeLa Cells
Small Interfering RNA
Amino Acid Transport Systems
Amino Acids
Two-Hybrid System Techniques
Hep G2 Cells
Hemagglutinins

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cell Biology

これを引用

Ohno, H., Nakatsu, Y., Sakoda, H., Kushiyama, A., Ono, H., Fujishiro, M., ... Asano, T. (2011). 4F2hc stabilizes GLUT1 protein and increases glucose transport activity. American Journal of Physiology - Cell Physiology, 300(5), C1047-C1054. https://doi.org/10.1152/ajpcell.00416.2010

4F2hc stabilizes GLUT1 protein and increases glucose transport activity. / Ohno, Haruya; Nakatsu, Yusuke; Sakoda, Hideyuki; Kushiyama, Akifumi; Ono, Hiraku; Fujishiro, Midori; Otani, Yuichiro; Okubo, Hirofumi; Yoneda, Masayasu; Fukushima, Toshiaki; Tsuchiya, Yoshihiro; Kamata, Hideaki; Nishimura, Fusanori; Kurihara, Hiroki; Katagiri, Hideki; Oka, Yoshitomo; Asano, Tomoichiro.

:: American Journal of Physiology - Cell Physiology, 巻 300, 番号 5, 01.05.2011, p. C1047-C1054.

研究成果: ジャーナルへの寄稿記事

Ohno, H, Nakatsu, Y, Sakoda, H, Kushiyama, A, Ono, H, Fujishiro, M, Otani, Y, Okubo, H, Yoneda, M, Fukushima, T, Tsuchiya, Y, Kamata, H, Nishimura, F, Kurihara, H, Katagiri, H, Oka, Y & Asano, T 2011, '4F2hc stabilizes GLUT1 protein and increases glucose transport activity', American Journal of Physiology - Cell Physiology, 巻. 300, 番号 5, pp. C1047-C1054. https://doi.org/10.1152/ajpcell.00416.2010
Ohno, Haruya ; Nakatsu, Yusuke ; Sakoda, Hideyuki ; Kushiyama, Akifumi ; Ono, Hiraku ; Fujishiro, Midori ; Otani, Yuichiro ; Okubo, Hirofumi ; Yoneda, Masayasu ; Fukushima, Toshiaki ; Tsuchiya, Yoshihiro ; Kamata, Hideaki ; Nishimura, Fusanori ; Kurihara, Hiroki ; Katagiri, Hideki ; Oka, Yoshitomo ; Asano, Tomoichiro. / 4F2hc stabilizes GLUT1 protein and increases glucose transport activity. :: American Journal of Physiology - Cell Physiology. 2011 ; 巻 300, 番号 5. pp. C1047-C1054.
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abstract = "Glucose transporter 1 (GLUT1) is widely distributed throughout various tissues and contributes to insulin- independent basal glucose uptake. Using a split-ubiquitin membrane yeast two-hybrid system, we newly identified 4F2 heavy chain (4F2hc) as a membrane protein interacting with GLUT1. Though 4F2hc reportedly forms heterodimeric complexes between amino acid transporters, such as LAT1 and LAT2, and regulates amino acid uptake, we investigated the effects of 4F2hc on GLUT1 expression and the associated glucose uptake. First, FLAG-tagged 4F2hc and hemagglutinin-tagged GLUT1 were overexpressed in human embryonic kidney 293 cells and their association was confirmed by coimmunoprecipitation. The green fluorescent protein-tagged 4F2hc and DsRed-tagged GLUT1 showed significant, but incomplete, colocalization at the plasma membrane. In addition, an endogenous association between GLUT1 and 4F2hc was demonstrated using mouse brain tissue and HeLa cells. Interestingly, overexpression of 4F2hc increased the amount of GLUT1 protein in HeLa and HepG2 cells with increased glucose uptake. In contrast, small interfering RNA (siRNA)-mediated 4F2hc gene suppression markedly reduced GLUT1 protein in both cell types, with reduced glucose uptake. While GLUT1 mRNA levels were not affected by overexpression or gene silencing of 4F2hc, GLUT1 degradation after the addition of cycloheximide was significantly suppressed by 4F2hc overexpression and increased by 4F2hc siRNA treatment. Taken together, these observations indicate that 4F2hc is likely to be involved in GLUT1 stabilization and to contribute to the regulation of not only amino acid but also glucose metabolism.",
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AU - Ohno, Haruya

AU - Nakatsu, Yusuke

AU - Sakoda, Hideyuki

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AU - Ono, Hiraku

AU - Fujishiro, Midori

AU - Otani, Yuichiro

AU - Okubo, Hirofumi

AU - Yoneda, Masayasu

AU - Fukushima, Toshiaki

AU - Tsuchiya, Yoshihiro

AU - Kamata, Hideaki

AU - Nishimura, Fusanori

AU - Kurihara, Hiroki

AU - Katagiri, Hideki

AU - Oka, Yoshitomo

AU - Asano, Tomoichiro

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N2 - Glucose transporter 1 (GLUT1) is widely distributed throughout various tissues and contributes to insulin- independent basal glucose uptake. Using a split-ubiquitin membrane yeast two-hybrid system, we newly identified 4F2 heavy chain (4F2hc) as a membrane protein interacting with GLUT1. Though 4F2hc reportedly forms heterodimeric complexes between amino acid transporters, such as LAT1 and LAT2, and regulates amino acid uptake, we investigated the effects of 4F2hc on GLUT1 expression and the associated glucose uptake. First, FLAG-tagged 4F2hc and hemagglutinin-tagged GLUT1 were overexpressed in human embryonic kidney 293 cells and their association was confirmed by coimmunoprecipitation. The green fluorescent protein-tagged 4F2hc and DsRed-tagged GLUT1 showed significant, but incomplete, colocalization at the plasma membrane. In addition, an endogenous association between GLUT1 and 4F2hc was demonstrated using mouse brain tissue and HeLa cells. Interestingly, overexpression of 4F2hc increased the amount of GLUT1 protein in HeLa and HepG2 cells with increased glucose uptake. In contrast, small interfering RNA (siRNA)-mediated 4F2hc gene suppression markedly reduced GLUT1 protein in both cell types, with reduced glucose uptake. While GLUT1 mRNA levels were not affected by overexpression or gene silencing of 4F2hc, GLUT1 degradation after the addition of cycloheximide was significantly suppressed by 4F2hc overexpression and increased by 4F2hc siRNA treatment. Taken together, these observations indicate that 4F2hc is likely to be involved in GLUT1 stabilization and to contribute to the regulation of not only amino acid but also glucose metabolism.

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