Journal of Southern Medical University ›› 2013, Vol. 33 ›› Issue (03): 351-.

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Functional role of protein kinase D1 in Aspergillums fumigates-induced activation of nuclear factor-κB signal pathway and transcription

  

  • Online:2013-03-20 Published:2013-03-20

Abstract: Objective To explore the functional role of protein kinase D1 (PKD1) in the activation of nuclear factor-κB (NF-κB)
signal pathway and NF-κB transcription mediated by Aspergillums fumigates. Methods A549 cells and HEK293 cells were
transfected with green fluorescence protein (GFP) or GFP-PKD1 followed by treatment with 1×105 CFU/ml Aspergillus fumigatus
conidia for different time lengths. The phosphorylation levels of PKD1, IКB and p65 (pS276) in the transfected cells were
measured by Western blotting. A549 cells were transfected with GFP-PKD1 or siRNA-PKD1, and the phosphorylation of IКB
and p65 (pS276) was examined. Finally, NF-κB-luc and renilla luciferase reporter pRL-SV40 were cotransfected into GFP- or
GFP-PKD1-transfected A549 cells before exposure of the cells to Aspergillus fumigatus conidia for 24 h, and NF-κB
transcriptional activity in the cells was determined using dual-luciferase reporter assay. Results Overexpression of PKD1
significantly increased Aspergillus fumigatus conidia-stimulated phosphorylation of PKD1, IКB and p65 (pS276), whereas PKD1
knockdown by siRNA-PKD1 suppressed IКB and p65 (pS276) phosphorylation. Dual luciferase assay demonstrated that PKD1
overexpression markedly enhanced Aspergillus fumigatus-induced NF-κB transcription in A549 cells. Conclusion PKD1 may
contribute to the activation of NF-κB signal pathway and NF-κB transcription induced by Aspergillums fumigates.