Activation of Protein Kinase D by Signaling through Rho and the alpha Subunit of the Heterotrimeric G Protein G sub(13)

Protein kinase D (PKD/PKC mu ) immunoprecipitated from COS-7 cells transiently transfected with either a constitutively active mutant of Rho (RhoQ63L) or the Rho-specific guanine nucleotide exchange factor pOnco-Lbc (Lbc) exhibited a marked increase in basal activity. Addition of aluminum fluoride t...

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Bibliographic Details
Published in:The Journal of biological chemistry 2001-10, Vol.276 (42), p.38619-38627
Main Authors: Yuan, J, Slice, L W, Rozengurt, E
Format: Article
Language:English
Subjects:
aluminum fluoride
bombesin receptors
C3 toxin
Clostridium botulinum
guanine nucleotide exchange factor
protein kinase D
Rho protein
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Summary:Protein kinase D (PKD/PKC mu ) immunoprecipitated from COS-7 cells transiently transfected with either a constitutively active mutant of Rho (RhoQ63L) or the Rho-specific guanine nucleotide exchange factor pOnco-Lbc (Lbc) exhibited a marked increase in basal activity. Addition of aluminum fluoride to cells co-transfected with PKD and wild type G alpha sub(13) also induced PKD activation. Co-transfection of Clostridium botulinum C3 toxin blocked activation of PKD by RhoQ63L, Lbc, or aluminum fluoride-stimulated G alpha sub(13). Treatment with the protein kinase C inhibitors GF I or Ro 31-8220 prevented the increase in PKD activity induced by RhoQ63L, Lbc, or aluminum fluoride-stimulated G alpha sub(13). PKD activation in response to G alpha sub(13) signaling was also completely prevented by mutation of Ser-744 and Ser-748 to Ala in the kinase activation loop of PKD. Co-expression of C. botulinum C3 toxin and a COOH-terminal fragment of G alpha sub(q) that acts in a dominant-negative fashion blocked PKD activation in response to agonist stimulation of bombesin receptor. Expression of the COOH-terminal region of G alpha sub(13) also attenuated PKD activation in response to bombesin receptor stimulation. Our results show that G alpha sub(13) contributes to PKD activation through a Rho- and protein kinase C-dependent signaling pathway and indicate that PKD activation is mediated by both G alpha sub(q) and G alpha sub(13) in response to bombesin receptor stimulation.
ISSN:0021-9258