Monitoring NF-??B Transactivation Potential Via Real-Time PCR Quantification of I??B-?? Gene Expression

Genetic pathologies involving deficits in DNA repair, like xeroderma pigmentosum (XP), show recurrent cell death, tissue degeneration and oncogenesis due to high sensitivity to ultraviolet radiation (UV). Various inducers including UV activate NF-κB, a pathway largely involved in cell proliferation and apoptosis. However, the mechanism(s) involving NF-κB activation by UV are poorly understood. To improve this knowledge, we examined NF-κB in two XP cell groups (XPC and XPD/TTD). XPC/D primary fibroblasts possess functional NF-κB dimers, and pro-inflammatory cytokines consistently activate NF-κB pathway. Contrarily, UV-mediated NF-κB activation is practically absent, whereas κB-specific DNA binding and transcriptional activity are dramatically undermined. These results indicate that lack of UV responsiveness at the NF-κB level is a common feature of XPC/D cells, suggesting that XP proteins might act upstream on NF-κB activity induced by UV. These observations help us to better understand the UV sensitivity and compromised survival of XP deficient cells.

show abstract

“…2A) and, concomitantly, we examined by Real-Time PCR the expression of the prototype NF-κB target gene IκBα 27 (Fig. 2B).…”

Section: Resultsmentioning

confidence: 99%

UV-Mediated NF-κB Activation is Abolished in Deficient XPC/D Primary Fibroblasts

Muotri¹,

Bottero²,

Tergaonkar³

et al. 2006

Cell Cycle

Self Cite

View full text Add to dashboard Cite

show abstract

“…Unfortunately, this effect was not detected at 3 h, neither by cDNA microarray (Table 2) nor by sqRT-PCR (Table 3). Since Bottero et al (7) have shown that activation of the NF-B protein complex closely correlates with the transcriptional level of NF-B IA, the measurement of its mRNA level appears as a rapid, sensitive, and powerful method to quantify the transcriptional power of NF-B. The increased expression of NF-B IA observed in our experiments therefore signed the activation of the NF-B complex in the bacterial supernatant condition and to a lesser extent in the live bacteria condition.…”

Section: Discussionmentioning

confidence: 99%

LiveStaphylococcus aureusand bacterial soluble factors induce different transcriptional responses in human airway cells

Moreilhon

Gras

Hologne

et al. 2005

Physiological Genomics

View full text Add to dashboard Cite

show abstract

“…The genes encoding nfkbia, nfkbiz, and rela were upregulated in cells treated with Sa and Sp, respectively. Bottero et al (10) have shown that the transcriptional level of nfkbia correlates well with the activation of the NF-B protein complex. The genes encoding NFB-REL proteins have previously been shown to be regulated after infections by diverse bacteria (7,24,30,34,60).…”

Section: Mec Responses To Live S Aureus or Bacterial Supernatantmentioning

confidence: 99%

Genetic susceptibility to S. aureus mastitis in sheep: differential expression of mammary epithelial cells in response to live bacteria or supernatant

Bonnefont

Rainard

Cunha

et al. 2012

Physiological Genomics

View full text Add to dashboard Cite

Staphylococcus aureus is a prevalent pathogen for mastitis in dairy ruminants and is responsible for both clinical and subclinical mastitis. Mammary epithelial cells (MEC) represent not only a physical barrier against bacterial invasion but are also active players of the innate immune response permitting infection clearance. To decipher their functions in general and in animals showing different levels of genetic predisposition to Staphylococcus in particular, MEC from ewes undergoing a divergent selection on milk somatic cell count were stimulated by S. aureus . MEC response was also studied according to the stimulation condition with live bacteria or culture supernatant. The early MEC response was studied during a 5 h time course by microarray to identify differentially expressed genes with regard to the host genetic background and as a function of the conditions of stimulation. In both conditions of stimulation, metabolic processes were altered, the apoptosis-associated pathways were considerably modified, and inflammatory and immune responses were enhanced with the upregulation of il1a, il1b, and tnfa and several chemokines known to enhance neutrophil ( cxcl8) or mononuclear leukocyte ( ccl20) recruitment. Genes associated with oxidative stress were increased after live bacteria stimulation, whereas immune response-related genes were higher after supernatant stimulation in the early phase. Only 20 genes were differentially expressed between Staphylococcus spp-mastitis resistant and susceptible animals without any clearly defined role on the control of infection. To conclude, this suggests that MEC may not represent the cell type at the origin of the difference of mastitis susceptibility, at least as demonstrated in our genetic model. Supernatant or heat-killed S. aureus produce biological effects that are essentially different from those induced by live bacteria.

show abstract

Monitoring NF-??B Transactivation Potential Via Real-Time PCR Quantification of I??B-?? Gene Expression

Cited by 33 publications

References 11 publications

UV-Mediated NF-κB Activation is Abolished in Deficient XPC/D Primary Fibroblasts

UV-Mediated NF-κB Activation is Abolished in Deficient XPC/D Primary Fibroblasts

LiveStaphylococcus aureusand bacterial soluble factors induce different transcriptional responses in human airway cells

Genetic susceptibility to S. aureus mastitis in sheep: differential expression of mammary epithelial cells in response to live bacteria or supernatant

Contact Info

Product

Resources

About

Monitoring NF-??B Transactivation Potential Via Real-Time PCR Quantification of I??B-?? Gene Expression

Cited by 33 publications

References 11 publications

UV-Mediated NF-&kappa;B Activation is Abolished in Deficient XPC/D Primary Fibroblasts

UV-Mediated NF-&kappa;B Activation is Abolished in Deficient XPC/D Primary Fibroblasts

LiveStaphylococcus aureusand bacterial soluble factors induce different transcriptional responses in human airway cells

Genetic susceptibility to S. aureus mastitis in sheep: differential expression of mammary epithelial cells in response to live bacteria or supernatant

Contact Info

Product

Resources

About

UV-Mediated NF-κB Activation is Abolished in Deficient XPC/D Primary Fibroblasts

UV-Mediated NF-κB Activation is Abolished in Deficient XPC/D Primary Fibroblasts