Analysis of insulin signalling by RNAi-based gene silencing

Q. I. Zhou, Jin Park, Z. Y. Jiang, J. J. Holik, P. Mitra, S. Semiz, A. Guilherme, A. M. Powelka, X. Tang, J. Virbasius, M. P. Czech

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Using siRNA-mediated gene silencing in cultured adipocytes, we have dissected the insulin-signalling pathway leading to translocation of GLUT4 glucose transporters to the plasma membrane. RNAi (RNA interference)-based depletion of components in the putative TC10 pathway (CAP, CrkII and c-Cbl plus Cbl-b) or the phospholipase Cγ pathway failed to diminish insulin signalling to GLUT4. Within the phosphoinositide 3-kinase pathway, loss of the 5′-phosphatidylinositol 3,4,5-trisphosphate phosphatase SHIP2 was also without effect, whereas depletion of the 3′-phosphatase PTEN significantly enhanced insulin action. Downstream of phosphatidylinositol 3,4,5-trisphosphate and PDK1, silencing the genes encoding the protein kinases Akt1/PKBα, or CISK(SGK3) or protein kinases Cλ/ζ had little or no effect, but loss of Akt2/PKBβ significantly attenuated GLUT4 regulation by insulin. These results show that Akt2/PKBβ is the key downstream intermediate within the phosphoinositide 3-kinase pathway linked to insulin action on GLUT4 in cultured adipocytes, whereas PTEN is a potent negative regulator of this pathway.

Original languageEnglish (US)
Pages (from-to)817-821
Number of pages5
JournalBiochemical Society Transactions
Volume32
Issue number5
DOIs
StatePublished - Nov 1 2004
Externally publishedYes

Fingerprint

Gene Silencing
RNA Interference
Genes
RNA
Insulin
1-Phosphatidylinositol 4-Kinase
Phosphatidylinositols
Adipocytes
Phosphotransferases
PTEN Phosphohydrolase
Gene encoding
Facilitative Glucose Transport Proteins
Type C Phospholipases
Cell membranes
Protein Kinases
Protein Kinase C
Small Interfering RNA
Cell Membrane
Proteins

Keywords

  • Gene silencing
  • Glucose
  • Insulin signalling
  • Phosphoinositide 3-kinase (PI3K)
  • Phospholipase Cγ
  • Protien kinase Cλ/ζ siRNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Zhou, Q. I., Park, J., Jiang, Z. Y., Holik, J. J., Mitra, P., Semiz, S., ... Czech, M. P. (2004). Analysis of insulin signalling by RNAi-based gene silencing. Biochemical Society Transactions, 32(5), 817-821. https://doi.org/10.1042/BST0320817

Analysis of insulin signalling by RNAi-based gene silencing. / Zhou, Q. I.; Park, Jin; Jiang, Z. Y.; Holik, J. J.; Mitra, P.; Semiz, S.; Guilherme, A.; Powelka, A. M.; Tang, X.; Virbasius, J.; Czech, M. P.

In: Biochemical Society Transactions, Vol. 32, No. 5, 01.11.2004, p. 817-821.

Research output: Contribution to journalArticle

Zhou, QI, Park, J, Jiang, ZY, Holik, JJ, Mitra, P, Semiz, S, Guilherme, A, Powelka, AM, Tang, X, Virbasius, J & Czech, MP 2004, 'Analysis of insulin signalling by RNAi-based gene silencing', Biochemical Society Transactions, vol. 32, no. 5, pp. 817-821. https://doi.org/10.1042/BST0320817
Zhou, Q. I. ; Park, Jin ; Jiang, Z. Y. ; Holik, J. J. ; Mitra, P. ; Semiz, S. ; Guilherme, A. ; Powelka, A. M. ; Tang, X. ; Virbasius, J. ; Czech, M. P. / Analysis of insulin signalling by RNAi-based gene silencing. In: Biochemical Society Transactions. 2004 ; Vol. 32, No. 5. pp. 817-821.
@article{7889a6471d3a4f9fb11778be40252ac4,
title = "Analysis of insulin signalling by RNAi-based gene silencing",
abstract = "Using siRNA-mediated gene silencing in cultured adipocytes, we have dissected the insulin-signalling pathway leading to translocation of GLUT4 glucose transporters to the plasma membrane. RNAi (RNA interference)-based depletion of components in the putative TC10 pathway (CAP, CrkII and c-Cbl plus Cbl-b) or the phospholipase Cγ pathway failed to diminish insulin signalling to GLUT4. Within the phosphoinositide 3-kinase pathway, loss of the 5′-phosphatidylinositol 3,4,5-trisphosphate phosphatase SHIP2 was also without effect, whereas depletion of the 3′-phosphatase PTEN significantly enhanced insulin action. Downstream of phosphatidylinositol 3,4,5-trisphosphate and PDK1, silencing the genes encoding the protein kinases Akt1/PKBα, or CISK(SGK3) or protein kinases Cλ/ζ had little or no effect, but loss of Akt2/PKBβ significantly attenuated GLUT4 regulation by insulin. These results show that Akt2/PKBβ is the key downstream intermediate within the phosphoinositide 3-kinase pathway linked to insulin action on GLUT4 in cultured adipocytes, whereas PTEN is a potent negative regulator of this pathway.",
keywords = "Gene silencing, Glucose, Insulin signalling, Phosphoinositide 3-kinase (PI3K), Phospholipase Cγ, Protien kinase Cλ/ζ siRNA",
author = "Zhou, {Q. I.} and Jin Park and Jiang, {Z. Y.} and Holik, {J. J.} and P. Mitra and S. Semiz and A. Guilherme and Powelka, {A. M.} and X. Tang and J. Virbasius and Czech, {M. P.}",
year = "2004",
month = "11",
day = "1",
doi = "10.1042/BST0320817",
language = "English (US)",
volume = "32",
pages = "817--821",
journal = "Biochemical Society Transactions",
issn = "0300-5127",
publisher = "Portland Press Ltd.",
number = "5",

}

TY - JOUR

T1 - Analysis of insulin signalling by RNAi-based gene silencing

AU - Zhou, Q. I.

AU - Park, Jin

AU - Jiang, Z. Y.

AU - Holik, J. J.

AU - Mitra, P.

AU - Semiz, S.

AU - Guilherme, A.

AU - Powelka, A. M.

AU - Tang, X.

AU - Virbasius, J.

AU - Czech, M. P.

PY - 2004/11/1

Y1 - 2004/11/1

N2 - Using siRNA-mediated gene silencing in cultured adipocytes, we have dissected the insulin-signalling pathway leading to translocation of GLUT4 glucose transporters to the plasma membrane. RNAi (RNA interference)-based depletion of components in the putative TC10 pathway (CAP, CrkII and c-Cbl plus Cbl-b) or the phospholipase Cγ pathway failed to diminish insulin signalling to GLUT4. Within the phosphoinositide 3-kinase pathway, loss of the 5′-phosphatidylinositol 3,4,5-trisphosphate phosphatase SHIP2 was also without effect, whereas depletion of the 3′-phosphatase PTEN significantly enhanced insulin action. Downstream of phosphatidylinositol 3,4,5-trisphosphate and PDK1, silencing the genes encoding the protein kinases Akt1/PKBα, or CISK(SGK3) or protein kinases Cλ/ζ had little or no effect, but loss of Akt2/PKBβ significantly attenuated GLUT4 regulation by insulin. These results show that Akt2/PKBβ is the key downstream intermediate within the phosphoinositide 3-kinase pathway linked to insulin action on GLUT4 in cultured adipocytes, whereas PTEN is a potent negative regulator of this pathway.

AB - Using siRNA-mediated gene silencing in cultured adipocytes, we have dissected the insulin-signalling pathway leading to translocation of GLUT4 glucose transporters to the plasma membrane. RNAi (RNA interference)-based depletion of components in the putative TC10 pathway (CAP, CrkII and c-Cbl plus Cbl-b) or the phospholipase Cγ pathway failed to diminish insulin signalling to GLUT4. Within the phosphoinositide 3-kinase pathway, loss of the 5′-phosphatidylinositol 3,4,5-trisphosphate phosphatase SHIP2 was also without effect, whereas depletion of the 3′-phosphatase PTEN significantly enhanced insulin action. Downstream of phosphatidylinositol 3,4,5-trisphosphate and PDK1, silencing the genes encoding the protein kinases Akt1/PKBα, or CISK(SGK3) or protein kinases Cλ/ζ had little or no effect, but loss of Akt2/PKBβ significantly attenuated GLUT4 regulation by insulin. These results show that Akt2/PKBβ is the key downstream intermediate within the phosphoinositide 3-kinase pathway linked to insulin action on GLUT4 in cultured adipocytes, whereas PTEN is a potent negative regulator of this pathway.

KW - Gene silencing

KW - Glucose

KW - Insulin signalling

KW - Phosphoinositide 3-kinase (PI3K)

KW - Phospholipase Cγ

KW - Protien kinase Cλ/ζ siRNA

UR - http://www.scopus.com/inward/record.url?scp=8744305112&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=8744305112&partnerID=8YFLogxK

U2 - 10.1042/BST0320817

DO - 10.1042/BST0320817

M3 - Article

C2 - 15494023

AN - SCOPUS:8744305112

VL - 32

SP - 817

EP - 821

JO - Biochemical Society Transactions

JF - Biochemical Society Transactions

SN - 0300-5127

IS - 5

ER -