Does anybody know what the phosphorylation of different Serine residue does to NFkB p65 activation?
I have an assay by Western blot detecting pS536 of p65 and a flow cytometry assay using pS529 of p65. I don't know if both would reflect the same thing.
Ser 536 is phosphorylated by IKKbeta while Ser 529 by CKII. In TNFalpha induction pSer536 but not 529 is required for p65 activation at least in some cell systems.
Ser 536 is phosphorylated by IKKbeta while Ser 529 by CKII. In TNFalpha induction pSer536 but not 529 is required for p65 activation at least in some cell systems.
Both phosphorylation sites regulate positively the activity of p65. But are involved also in different activities. p-S536 is implicated in apoptosis. For a complete p-sites and implications look at
http://www.phosphosite.org/proteinAction.do?id=1012&showAllSites=true
Hello Sylviane,
did you try phosphosite.org (http://www.phosphosite.org/proteinAction.do?id=1012&showAllSites=true)? There you will find a list of phosphorylation sites and their implication/function.
Good luck :)
Yes, Federico Galvagni is right. These two sites are phosphorylated by two different upstream kinases. In my own opinion, IKK-mediated S536 phosphorylation is critical for activation of the canonical NF-kB pathway, while S529 may increase NF-kB activity. In our research, we found that IKK-dependent S536 phosphorylation is also required for RelA/p65 nulcear translocation, acetylation in nucleus, and thus NF-kB activation induced by HDAC inhibitors (J Biol Chem. 2011 Sep 30;286(39):34036-50. doi: 10.1074/jbc.M111.284216. Epub 2011 Aug 4.
Disruption of IkappaB kinase (IKK)-mediated RelA serine 536 phosphorylation sensitizes human multiple myeloma cells to histone deacetylase (HDAC) inhibitors.)
Thank you All, that's very helpful and that explains the discrepancy between my Western and Flow results. Thanks again.
Dear sylviane
The phosphorylation of p60 in two serine site, is necessary for full activation of NFkB. After phosphorylation, the conformation structure of p60 completely suitable for attach to p50.
Dear Sylvane,
Following what the previous colleagues have mentioned, this review summarizes all the possible NF-κB subunit phosphorylations. Just in case you want to go deeper into the subject:
Article The Regulation of NF-κB Subunits by Phosphorylation
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