The phosphatidic acid-binding, polybasic domain is responsible for the differences in the phosphoregulation of lipins 1 and 3.

Salome Boroda, Sankeerth Takkellapati, Robert T Lawrence, Samuel W Entwisle, Jennifer M Pearson, Mitchell E Granade, Garrett R Mullins, James M Eaton, Judit Villén, Thurl E Harris

The Journal of biological chemistry 2017 Dec 15

filter terms:

Lipins 1, 2, and 3 are Mg2+-dependent phosphatidic acid phosphatases and catalyze the penultimate step of triacylglycerol synthesis. We have previously investigated the biochemistry of lipins 1 and 2 and shown that di-anionic phosphatidic acid (PA) augments their activity and lipid binding and that lipin 1 activity is negatively regulated by phosphorylation. In the present study, we show that phosphorylation does not affect the catalytic activity of lipin 3 or its ability to associate with PA in vitro The lipin proteins each contain a conserved polybasic domain (PBD) composed of nine lysine and arginine residues located between the conserved N- and C-terminal domains. In lipin 1, the PBD is the site of PA binding and sensing of the PA electrostatic charge. The specific arrangement and number of the lysines and arginines of the PBD vary among the lipins. We show that the different PBDs of lipins 1 and 3 are responsible for the presence of phosphoregulation on the former but not the latter enzyme. To do so, we generated lipin 1 that contained the PBD of lipin 3 and vice versa. The lipin 1 enzyme with the lipin 3 PBD lost its ability to be regulated by phosphorylation but remained downstream of phosphorylation by mammalian target of rapamycin. Conversely, the presence of the lipin 1 PBD in lipin 3 subjected the enzyme to negative intramolecular control by phosphorylation. These results indicate a mechanism for the observed differences in lipin phosphoregulation in vitro. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Citation

Salome Boroda, Sankeerth Takkellapati, Robert T Lawrence, Samuel W Entwisle, Jennifer M Pearson, Mitchell E Granade, Garrett R Mullins, James M Eaton, Judit Villén, Thurl E Harris. The phosphatidic acid-binding, polybasic domain is responsible for the differences in the phosphoregulation of lipins 1 and 3. The Journal of biological chemistry. 2017 Dec 15;292(50):20481-20493