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Learn MorePseudomonas aeruginosa use quorum-sensing molecules, including N-(3-oxododecanoyl)-homoserine lactone (C12), for intercellular communication. C12 activated apoptosis in mouse embryo fibroblasts (MEF) from both wild type (WT) and Bax/Bak double knock-out mice (WT MEF and DKO MEF that were responsive to C12, DKORMEF): nuclei fragmented; mitochondrial membrane potential (mito) depolarized; Ca2+was released from the endoplasmic reticulum (ER), increasing cytosolic [Ca2+] (Cacyto); caspase 3/7 was activated. DKORMEF had been isolated from a nonclonal pool of DKO MEF that were non-responsive to C12 (DKONRMEF). RNAseq analysis, qPCR and western blots showed that WT and DKORMEF both expressed genes associated with cancer, including paraoxonase 2 (PON2), while DKONRMEF expressed little PON2. Adenovirus-mediated expression of human PON2 in DKONRMEF rendered them responsive to C12: mitodepolarized, Cacytoincreased and caspase 3/7 activated. Human embryonic kidney 293T (HEK293T) cells expressed low levels of endogenous PON2, and these cells were also less responsive to C12. Overexpression of PON2, but not PON2-H114Q (no lactonase activity) in HEK293T cells caused them to become sensitive to C12. Because [C12] may reach high levels in biofilms in lungs of cystic fibrosis (CF) patients, PON2 lactonase activity may control mito, Ca2+release from the ER and apoptosis in CF airway epithelia. Coupled with previous data, these results also indicate that PON2 uses its lactonase activity to prevent Bax- and Bak-dependent apoptosis in response to common proapoptotic drugs like doxorubicin, staurosporine but activates Bax- and Bak-independent apoptosis in response to C12. SOURCE: Terry,E,Machen (tmachen@berkeley.edu) - Univ California-Berkeley
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