title: A GEF-to-phospholipase molecular switch caused by phosphatidic acid, Rac and JAK tyrosine kinase that explains leukocyte cell migration.
abstract: Phospholipase D2 (PLD2) is a cell-signaling molecule that bears two activities: a guanine-nucleotide exchange factor (GEF) and a lipase that reside in the PX/PH domains and in two HKD domains, respectively. Upon cell stimulation, the GEF activity yields Rac2-GTP and the lipase activity yields phosphatidic acid (PA). In the present study, we show for the first time that these activities regulate one another. Upon cell stimulation, both GEF and lipase activities are quickly (within ∼3 min) elevated. As soon as it is produced, PA positively feeds back on the GEF and further activates it. Rac2-GTP, on the other hand, is inhibitory to the lipase activity. PLD2 would remain downregulated if it were not for the contribution of the tyrosine kinase Janus kinase 3 (JAK3), which restores lipase action (by phosphorylation at Y415). Conversely, the GEF is inhibited upon phosphorylation by JAK3 and is effectively terminated by this action and by the increasing accumulation of PA at >15 min of cell stimulation. This PA interferes with the ability of the GEF to bind to its substrate (Rac2-GTP). Thus, both temporal inter-regulation and phosphorylation-dependent mechanisms are involved in determining a GEF-lipase switch within the same molecule. Human neutrophils stimulated by interleukin-8 follow a biphasic pattern of GEF and lipase activation that can be explained by such an intramolecular switch. This is the first report of a temporal inter-regulation of two enzymatic activities that reside in the same molecule with profound biological consequences in leukocyte cell migration.
authors: Department of Biochemistry and Molecular Biology, Wright State University School of Medicine, OH 45435, USA.
authors: Madhu
authors: M
authors: Mahankali
authors: Karen M
authors: KM
authors: Henkels
authors: Julian
authors: J
authors: Gomez-Cambronero
mesh_terms: D000818:Animals; D019556:COS Cells; D002465:Cell Movement / Q000235:genetics; D002522:Chlorocebus aethiops; D025461:Feedback, Physiological; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics; D006801:Humans; D053616:Janus Kinase 3 / Q000235:genetics / Q000378:metabolism*; D007962:Leukocytes / Q000502:physiology*; D010712:Phosphatidic Acids / Q000378:metabolism*; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D019076:Transgenes / Q000235:genetics; D020744:rac GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000097766:RAC2 GTP-Binding Protein
chemical_list: D020662:Guanine Nucleotide Exchange Factors; D053616:Janus Kinase 3; D010712:Phosphatidic Acids; D010739:Phospholipase D; D034741:RNA, Small Interfering; D020744:rac GTP-Binding Proteins; D000097766:RAC2 GTP-Binding Protein; C105606:phospholipase D2
keywords: Cell migration; Cell signalling; Chemotaxis; HL-60 cells; Leukocyte; Tyrosine kinase JAK
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