Cdc42-dependent Mediation of UV-induced p38 Activation by G Protein βγ Subunits

Mi Ran Seo, Chin Ho Cho, Yun Il Lee, Eun Young Shin, Dongeun Park, Chang Dae Bae, Jung Weon Lee, Eun So Lee, Yong-Sung Juhnn

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Abstract

The β and γ subunits of heterotrimeric GTP-binding proteins (Gβγ) were found to bi-directionally regulate the UV-induced activation of p38 and c-Jun NH2-terminal kinase, and the UV-induced activation of p38 was reported to enhance the resistance of normal keratinocytes to apoptosis. However, the signaling pathway downstream of Gβγ for this UV-induced p38 activation is not known. Thus, we examined the role of the Rho GTPase family in the regulation of UV-induced p38 activation by Gβγ. We found that overexpression of Gβγ increased the UV-induced activation of Cdc42 and that overexpression of constitutively active V12 Cdc42 increased the UV-induced p38 activation. Transfection of dominant negative N17 Cdc42 or small interfering RNA for Cdc42 blocked UV-induced p38 activation mediated by Gβγ in COS-1 and HaCaT cells. UV-induced p38 activation by Gβγ was blocked by overexpression of dominant negative p21-activated kinase (PAK)-interacting exchange factor β (βPix), and wild type βPix stimulated the UV-induced p38 activation, which was blocked by N17 Cdc42. Gβγ increased the UV-induced activation of Ras, and the overexpression of V12 Ras increased UV-induced p38 activation, which was blocked by dominant negative βPix. UV-induced p38 activation was inhibited by N17 Ras and a farnesyltransferase inhibitor, manumycin A. Gβγ also increased the UV-induced phosphorylation of the epidermal growth factor receptor (EGFR), and the UV-induced p38 activation was blocked by an EGFR kinase inhibitor, AG1478. From these results, we conclude that Gβγ mediates UV-induced activation of p38 in a Cdc42-dependent way and that EGFR, Ras, and βPix act sequentially upstream of Cdc42 in COS-1 and HaCaT cells.

Original languageEnglish
Pages (from-to)17366-17375
Number of pages10
JournalJournal of Biological Chemistry
Volume279
Issue number17
DOIs
StatePublished - 23 Apr 2004

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Protein Subunits
GTP-Binding Proteins
Epidermal Growth Factor Receptor
COS Cells
Chemical activation
p21-Activated Kinases
Farnesyltranstransferase
Heterotrimeric GTP-Binding Proteins
rho GTP-Binding Proteins
JNK Mitogen-Activated Protein Kinases
Keratinocytes
Small Interfering RNA
Transfection
Phosphorylation
Apoptosis

Cite this

Seo, Mi Ran ; Cho, Chin Ho ; Lee, Yun Il ; Shin, Eun Young ; Park, Dongeun ; Bae, Chang Dae ; Lee, Jung Weon ; Lee, Eun So ; Juhnn, Yong-Sung. / Cdc42-dependent Mediation of UV-induced p38 Activation by G Protein βγ Subunits. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 17. pp. 17366-17375.
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title = "Cdc42-dependent Mediation of UV-induced p38 Activation by G Protein βγ Subunits",
abstract = "The β and γ subunits of heterotrimeric GTP-binding proteins (Gβγ) were found to bi-directionally regulate the UV-induced activation of p38 and c-Jun NH2-terminal kinase, and the UV-induced activation of p38 was reported to enhance the resistance of normal keratinocytes to apoptosis. However, the signaling pathway downstream of Gβγ for this UV-induced p38 activation is not known. Thus, we examined the role of the Rho GTPase family in the regulation of UV-induced p38 activation by Gβγ. We found that overexpression of Gβγ increased the UV-induced activation of Cdc42 and that overexpression of constitutively active V12 Cdc42 increased the UV-induced p38 activation. Transfection of dominant negative N17 Cdc42 or small interfering RNA for Cdc42 blocked UV-induced p38 activation mediated by Gβγ in COS-1 and HaCaT cells. UV-induced p38 activation by Gβγ was blocked by overexpression of dominant negative p21-activated kinase (PAK)-interacting exchange factor β (βPix), and wild type βPix stimulated the UV-induced p38 activation, which was blocked by N17 Cdc42. Gβγ increased the UV-induced activation of Ras, and the overexpression of V12 Ras increased UV-induced p38 activation, which was blocked by dominant negative βPix. UV-induced p38 activation was inhibited by N17 Ras and a farnesyltransferase inhibitor, manumycin A. Gβγ also increased the UV-induced phosphorylation of the epidermal growth factor receptor (EGFR), and the UV-induced p38 activation was blocked by an EGFR kinase inhibitor, AG1478. From these results, we conclude that Gβγ mediates UV-induced activation of p38 in a Cdc42-dependent way and that EGFR, Ras, and βPix act sequentially upstream of Cdc42 in COS-1 and HaCaT cells.",
author = "Seo, {Mi Ran} and Cho, {Chin Ho} and Lee, {Yun Il} and Shin, {Eun Young} and Dongeun Park and Bae, {Chang Dae} and Lee, {Jung Weon} and Lee, {Eun So} and Yong-Sung Juhnn",
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Seo, MR, Cho, CH, Lee, YI, Shin, EY, Park, D, Bae, CD, Lee, JW, Lee, ES & Juhnn, Y-S 2004, 'Cdc42-dependent Mediation of UV-induced p38 Activation by G Protein βγ Subunits', Journal of Biological Chemistry, vol. 279, no. 17, pp. 17366-17375. https://doi.org/10.1074/jbc.M312442200

Cdc42-dependent Mediation of UV-induced p38 Activation by G Protein βγ Subunits. / Seo, Mi Ran; Cho, Chin Ho; Lee, Yun Il; Shin, Eun Young; Park, Dongeun; Bae, Chang Dae; Lee, Jung Weon; Lee, Eun So; Juhnn, Yong-Sung.

In: Journal of Biological Chemistry, Vol. 279, No. 17, 23.04.2004, p. 17366-17375.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Cdc42-dependent Mediation of UV-induced p38 Activation by G Protein βγ Subunits

AU - Seo, Mi Ran

AU - Cho, Chin Ho

AU - Lee, Yun Il

AU - Shin, Eun Young

AU - Park, Dongeun

AU - Bae, Chang Dae

AU - Lee, Jung Weon

AU - Lee, Eun So

AU - Juhnn, Yong-Sung

PY - 2004/4/23

Y1 - 2004/4/23

N2 - The β and γ subunits of heterotrimeric GTP-binding proteins (Gβγ) were found to bi-directionally regulate the UV-induced activation of p38 and c-Jun NH2-terminal kinase, and the UV-induced activation of p38 was reported to enhance the resistance of normal keratinocytes to apoptosis. However, the signaling pathway downstream of Gβγ for this UV-induced p38 activation is not known. Thus, we examined the role of the Rho GTPase family in the regulation of UV-induced p38 activation by Gβγ. We found that overexpression of Gβγ increased the UV-induced activation of Cdc42 and that overexpression of constitutively active V12 Cdc42 increased the UV-induced p38 activation. Transfection of dominant negative N17 Cdc42 or small interfering RNA for Cdc42 blocked UV-induced p38 activation mediated by Gβγ in COS-1 and HaCaT cells. UV-induced p38 activation by Gβγ was blocked by overexpression of dominant negative p21-activated kinase (PAK)-interacting exchange factor β (βPix), and wild type βPix stimulated the UV-induced p38 activation, which was blocked by N17 Cdc42. Gβγ increased the UV-induced activation of Ras, and the overexpression of V12 Ras increased UV-induced p38 activation, which was blocked by dominant negative βPix. UV-induced p38 activation was inhibited by N17 Ras and a farnesyltransferase inhibitor, manumycin A. Gβγ also increased the UV-induced phosphorylation of the epidermal growth factor receptor (EGFR), and the UV-induced p38 activation was blocked by an EGFR kinase inhibitor, AG1478. From these results, we conclude that Gβγ mediates UV-induced activation of p38 in a Cdc42-dependent way and that EGFR, Ras, and βPix act sequentially upstream of Cdc42 in COS-1 and HaCaT cells.

AB - The β and γ subunits of heterotrimeric GTP-binding proteins (Gβγ) were found to bi-directionally regulate the UV-induced activation of p38 and c-Jun NH2-terminal kinase, and the UV-induced activation of p38 was reported to enhance the resistance of normal keratinocytes to apoptosis. However, the signaling pathway downstream of Gβγ for this UV-induced p38 activation is not known. Thus, we examined the role of the Rho GTPase family in the regulation of UV-induced p38 activation by Gβγ. We found that overexpression of Gβγ increased the UV-induced activation of Cdc42 and that overexpression of constitutively active V12 Cdc42 increased the UV-induced p38 activation. Transfection of dominant negative N17 Cdc42 or small interfering RNA for Cdc42 blocked UV-induced p38 activation mediated by Gβγ in COS-1 and HaCaT cells. UV-induced p38 activation by Gβγ was blocked by overexpression of dominant negative p21-activated kinase (PAK)-interacting exchange factor β (βPix), and wild type βPix stimulated the UV-induced p38 activation, which was blocked by N17 Cdc42. Gβγ increased the UV-induced activation of Ras, and the overexpression of V12 Ras increased UV-induced p38 activation, which was blocked by dominant negative βPix. UV-induced p38 activation was inhibited by N17 Ras and a farnesyltransferase inhibitor, manumycin A. Gβγ also increased the UV-induced phosphorylation of the epidermal growth factor receptor (EGFR), and the UV-induced p38 activation was blocked by an EGFR kinase inhibitor, AG1478. From these results, we conclude that Gβγ mediates UV-induced activation of p38 in a Cdc42-dependent way and that EGFR, Ras, and βPix act sequentially upstream of Cdc42 in COS-1 and HaCaT cells.

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DO - 10.1074/jbc.M312442200

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