| Summary: | Availability of free arachidonic acid (AA) constitutes a rate limiting factor for cellular eicosanoid synthesis. AA distributes differentially across membrane phospholipids, which is largely due to the action of coenzyme A-independent transacylase (CoA-IT), an enzyme that moves the fatty acid primarily from diacyl phospholipid species to ether-containing species, particularly the ethanolamine plasmalogens. In this work, we examined the dependence of AA remodeling on plasmalogen content using the murine macrophage cell line RAW264.7 and its plasmalogen-deficient variants RAW.12 and RAW.108. All three strains remodeled AA between phospholipids with similar magnitude and kinetics, thus demonstrating that cellular plasmalogen content does not influence the process. Cell stimulation with yeast-derived zymosan also had no effect on AA remodeling, but incubating the cells in AA-rich media markedly slowed down the process. Further, knockdown of cytosolic-group IVC phospholipase A<sub>2</sub>γ (cPLA<sub>2</sub>γ) by RNA silencing significantly reduced AA remodeling, while inhibition of other major phospholipase A<sub>2</sub> forms such as cytosolic phospholipase A<sub>2</sub>α, calcium-independent phospholipase A<sub>2</sub>β, or secreted phospholipase A<sub>2</sub> had no effect. These results uncover new regulatory features of CoA-IT-mediated transacylation reactions in cellular AA homeostasis and suggest a hitherto unrecognized role for cPLA<sub>2</sub>γ in maintaining membrane phospholipid composition via regulation of AA remodeling.
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