P100 Deficiency is insufficient for full activation of the alternative NF-?B pathway: TNF cooperates with p52-RelB in target gene transcription.

ABSTRACT: BACKGROUND: Constitutive activation of the alternative NF-?B pathway leads to marginal zone B cell expansion and disorganized spleen microarchitecture. Furthermore, uncontrolled alternative NF-?B signaling may result in the development and progression of cancer. Here, we focused on the question how does the constitutive alternative NF-?B signaling exert its effects in these malignant processes. METHODOLOGY/PRINCIPAL FINDINGS: To explore the consequences of unrestricted alternative NF-?B activation on genome-wide transcription, we compared gene expression profiles of wild-type and NF-?B2/p100-deficient (p100(-/-)) primary mouse embryonic fibroblasts (MEFs) and spleens. Microarray experiments revealed only 73 differentially regulated genes in p100(-/-) vs. wild-type MEFs. Chromatin immunoprecipitation (ChIP) assays showed in p100(-/-) MEFs direct binding of p52 and RelB to the promoter of the Enpp2 gene encoding ENPP2/Autotaxin, a protein with an important role in lymphocyte homing and cell migration. Gene ontology analysis revealed upregulation of genes with anti-apoptotic/proliferative activity (Enpp2/Atx, Serpina3g, Traf1, Rrad), chemotactic/locomotory activity (Enpp2/Atx, Ccl8), and lymphocyte homing activity (Enpp2/Atx, Cd34). Most importantly, biochemical and gene expression analyses of MEFs and spleen, respectively, indicated a marked crosstalk between classical and alternative NF-?B pathways. CONCLUSIONS/SIGNIFICANCE: Our results show that p100 deficiency alone was insufficient for full induction of genes regulated by the alternative NF-?B pathway. Moreover, alternative NF-?B signaling strongly synergized both in vitro and in vivo with classical NF-?B activation, thereby extending the number of genes under the control of the p100 inhibitor of the alternative NF-?B signaling pathway.

SUBMITTER: Lovas A 

PROVIDER: S-EPMC3412832 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature