Use of RNA Interference-mediated Gene Silencing and Adenoviral Overexpression to Elucidate the Roles of AKT/Protein Kinase B Isoforms in Insulin Actions

Katome, Takashi; Obata, Toshiyuki; Matsushima, Rie; Masuyama, Norihisa; Cantley, Lewis C.; Gotoh, Yukiko; Kishi, Kazuhiro; Shiota, Hiroshi; Ebina, Yousuke

doi:10.1074/jbc.m302094200

Cited by 212Metabolism, Diabetes, and CancerPI3K/AKT/mTOR signaling in cancerPancreatic function and diabetes

Use of RNA Interference-mediated Gene Silencing and Adenoviral Overexpression to Elucidate the Roles of AKT/Protein Kinase B Isoforms in Insulin Actions

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Katome, Takashi, Obata, Toshiyuki, Matsushima, Rie, Masuyama, Norihisa, Cantley, Lewis C., Gotoh, Yukiko, Kishi, Kazuhiro, Shiota, Hiroshi +1 more·2003·Journal of Biological Chemistry

DOI 10.1074/jbc.m302094200

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Abstract

Insulin plays a central role in the regulation of glucose homeostasis in part by stimulating glucose uptake and glycogen synthesis. The serine/threonine protein kinase Akt has been proposed to mediate insulin signaling in several processes. However, it is unclear whether Akt is involved in insulin-stimulated glucose uptake and which isoforms of Akt are responsible for each insulin action. We confirmed that expression of a constitutively active Akt, using an adenoviral expression vector, promoted translocation of glucose transporter 4 (GLUT4) to plasma membrane, 2-deoxyglucose (2-DG) uptake, and glycogen synthesis in both Chinese hamster ovary cells and 3T3-L1 adipocytes. Inhibition of Akt either by adenoviral expression of a dominant negative Akt or by the introduction of synthetic 21-mer short interference RNA against Akt markedly reduced insulin-stimulated GLUT4 translocation, 2-DG uptake, and glycogen synthesis. Experiments with isoform-specific short interference RNA revealed that Akt2, and Akt1 to a lesser extent, has an essential role in insulin-stimulated GLUT4 translocation and 2-DG uptake in both cell lines, whereas Akt1 and Akt2 contribute equally to insulin-stimulated glycogen synthesis. These data suggest a prerequisite role of Akt in insulin-stimulated glucose uptake and distinct functions among Akt isoforms.

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PKBβ seems to regulate insulin-stimulated glucose uptake.
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From Glucocorticoid-induced insulin resistance in skeletal muscles: defects in insulin signalling and the effects of a selective glycogen synthase kinase-3 inhibitor (2005) · cites this paper as [13]

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Authors: Katome, T., Obata, T., Matsushima, R., Masuyama, N., Cantley, L. C., Gotoh, Y., Kishi, K., Shiota, H., & Ebina, Y.
Affiliations: Tokushima University (JP)
Beth Israel Deaconess Medical Center (US)
Institute for Quantitative Biosciences, The University of Tokyo (JP)
Corresponding author: Ebina, Yousuke
ORCID: Cantley, Lewis C.
Gotoh, Yukiko
Venue: Journal of Biological Chemistry
Publisher: Elsevier BV
Published: 2003-07
Volume: 278
Issue: 30
Pages: 28312-28323
ISSN: 0021-9258 (ISSN-L)
DOI: 10.1074/jbc.m302094200
OpenAlex: W2037308821
Cited by: 212
References: 54
Citations by year: 2025: 3 · 2023: 2 · 2022: 5 · 2021: 3 · 2020: 4 · 2019: 7 · 2018: 3 · 2017: 2 · 2016: 6 · 2015: 8 · 2014: 9 · 2013: 8 · 2012: 6
FWCI: 9.33
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: cc-by
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
creativecommons.org/licenses/by/4.0/ (vor, 6306 d)
Locations: Journal of Biological Chemistry (publishedVersion, cc-by)
PubMed (publishedVersion)
Funders: National Institute of General Medical Sciences (GM 41890, R01GM041890)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolism, Diabetes, and Cancer
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › PI3K/AKT/mTOR signaling in cancer
Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
Keywords: Akt, insulin signaling, insulin-stimulated glucose uptake, GLUT4 translocation, glycogen synthesis, AKT1, AKT2, siRNA, adenoviral overexpression, dominant-negative Akt, 2-deoxyglucose uptake, adipocytes, glucose homeostasis
MeSH terms: Adenoviridae, Animals, Base Sequence, Brain, Deoxyglucose, Dose-Response Relationship, Drug, Genetic Techniques, Glycogen, Cricetinae, Insulin, Luciferases, Molecular Sequence Data, Monosaccharide Transport Proteins, Muscle Proteins, Plasmids, Precipitin Tests, Proto-Oncogene Proteins, Time Factors, Immunoblotting, Gene Library, CHO Cells, 3T3 Cells, Protein Serine-Threonine Kinases, COS Cells, Protein Isoforms, Gene Silencing, Protein Transport, RNA Interference, Proto-Oncogene Proteins c-akt, Glucose Transporter Type 4, Mice, Rats

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Statistics and arithmetic are recomputed, not judged.

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Use of RNA Interference-mediated Gene Silencing and Adenoviral Overexpression to Elucidate the Roles of AKT/Protein Kinase B Isoforms in Insulin Actions

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Integrity signals

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