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 Fer-1 the wild form Corby, but not the flaA mutant, induced nuclear translocation of NF-kB.

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fibre7orange



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Date d'inscription : 22/01/2013

MessageSujet: Fer-1 the wild form Corby, but not the flaA mutant, induced nuclear translocation of NF-kB.   Ven 22 Aoû - 5:45

NF-kB is commonly existing inside the cytoplasm in an inactive state and is bound to members in the I B inhibitor protein family, chiefly I Ba. In this complex, I Ba blocks <br />the full report the nuclear localization signal, therefore pre venting nuclear translocation. Translocation of NF B to the nucleus needs disruption in the cytoplasmic NF B,I Ba complex. <br /><br />To find out the part of I Ba phosphorylation and degradation in L. pneumo phila induced NF B translocation and activation, we investigated whether L. pneumophila induces phosphor ylation and degradation of I Ba. The latter two pro cesses have been examined by Western blot analysis applying antibodies against phosphorylated and <br />selleck inhibitor complete I Ba, respectively. Fig. 6D exhibits phosphorylation and degra dation of I Ba in Jurkat cells contaminated together with the wild sort Corby but not the flaA mutant for 1, 2 and four h. The I Ba phosphorylation became evident at one h and decreased thereafter. Consistent with this, Corby induced degradation of I Ba was observed at 1 h. NF B signaling happens either through the classical or choice pathway. Inside the classical pathway, NF B dimers, such as p50/p65, are maintained within the cytoplasm by interaction with I Ba. <br /><br />Whereas the classical NF B activation is I B kinase b and IKKg dependent and happens by way of I Ba phosphoryla tion and subsequent proteasomal degradation, the alter native pathway depends on IKKa homodimers and NF B inducing kinase and final results in regulated processing of your p100 precursor protein to p52 via phosphorylation and degradation of its I B terminus. Without a doubt, the wild kind Corby but not the flaA mutant induced phosphorylation of p65 and upstream kinase IKKb. Subsequent, we examined the alterna tive pathway, which includes the cleavage of NF B2/ p100 to p52. The degree of p52 protein greater in Jurkat cells infected using the wild kind Corby but not the flaA mutant, indicating that flagellin activates NF B by means of the alternative pathway. NF B signal is crucial for induction of IL eight expression by L. <br /><br />pneumophila To additional confirm the involvement of I Ba degrada tion, we transfected the cells with transdominant mutant of I Ba through which two essential serine residues expected for inducer mediated phosphorylation have been deleted. As witnessed in Fig. 6E, overexpression of mutant I Ba considerably inhibited the Corby induced IL 8 promoter acti vation. This observation implicates the involvement of I Ba phosphorylation and degradation in flagellin induced IL 8 expression. To deal with the mechanism of flagellin mediated IL 8 expression, we investigated the function of NIK and IKK in L. pneumophila induced IL 8 expression. Cotransfection with all the dominant adverse mutant kinds of NIK, IKKa, IKKb, and IKKg inhibited L. pneumophila induced IL eight expression. MyD88 is a universal adaptor for induction of cytokines by TLR2, TLR4, TLR5, TLR7, and TLR9. It's also needed for activation of NF B by these TLRs. Likewise, overexpression of the dominant damaging mutant type of MyD88 also inhibited L. pneumophila induced IL 8 expression. Taken with each other, these findings obviously show that L. pneumophila induces IL 8 expression through activation of flagellin dependent NF B signaling pathway. Mainly because activation on the IL 8 promoter by L. pneumo phila infection expected the activation of NF B, we blocked NF B activation with Bay 11 7082, an inhibitor of I Ba phosphorylation.
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Fer-1 the wild form Corby, but not the flaA mutant, induced nuclear translocation of NF-kB.
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