Casein kinase II–mediated phosphorylation of lipin 1 phosphatidate phosphatase at Ser-285 and Ser-287 regulates its interaction with 14-3-3 protein

Meagan Hennessy, Mitchell E. Granade, Azam Hassaninasab, Dana Wang, Joanna M. Kwiatek, Gil Soo Han, Thurl E. Harris, George Carman

Research output: Contribution to journalArticle

Abstract

The mammalian lipin 1 phosphatidate phosphatase is a key regulatory enzyme in lipid metabolism. By catalyzing phosphatidate dephosphorylation, which produces diacylglycerol, the enzyme plays a major role in the synthesis of triacylglycerol and membrane phospholipids. The importance of lipin 1 to lipid metabolism is exemplified by cellular defects and lipid-based diseases associated with its loss or overexpression. Phosphorylation of lipin 1 governs whether it is associated with the cytoplasm apart from its substrate or with the endoplasmic reticulum membrane where its enzyme reaction occurs. Lipin 1 is phosphorylated on multiple sites, but less than 10% of them are ascribed to a specific protein kinase. Here, we demonstrate that lipin 1 is a bona fide substrate for casein kinase II (CKII), a protein kinase that is essential to viability and cell cycle progression. Phosphoamino acid analysis and phosphopeptide mapping revealed that lipin 1 is phosphorylated by CKII on multiple serine and threonine residues, with the former being major sites. Mutational analysis of lipin 1 and its peptides indicated that Ser-285 and Ser-287 are both phosphorylated by CKII. Substitutions of Ser-285 and Ser-287 with nonphosphorylatable alanine attenuated the interaction of lipin 1 with 14-3-3 protein, a regulatory hub that facilitates the cytoplasmic localization of phosphorylated lipin 1. These findings advance our understanding of how phosphorylation of lipin 1 phosphatidate phosphatase regulates its interaction with 14-3-3 protein and intracellular localization and uncover a mechanism by which CKII regulates cellular physiology.

Original languageEnglish (US)
Pages (from-to)2365-2374
Number of pages10
JournalJournal of Biological Chemistry
Volume294
Issue number7
DOIs
StatePublished - Jan 1 2019

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Casein Kinases
Phosphatidate Phosphatase
14-3-3 Proteins
Phosphorylation
Casein Kinase II
Lipid Metabolism
Protein Kinases
Enzymes
lipine
Phosphoamino Acids
Membranes
Phosphopeptides
Diglycerides
Physiology
Substrates
Threonine
Endoplasmic Reticulum
Alanine
Serine
Phospholipids

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Hennessy, Meagan ; Granade, Mitchell E. ; Hassaninasab, Azam ; Wang, Dana ; Kwiatek, Joanna M. ; Han, Gil Soo ; Harris, Thurl E. ; Carman, George. / Casein kinase II–mediated phosphorylation of lipin 1 phosphatidate phosphatase at Ser-285 and Ser-287 regulates its interaction with 14-3-3 protein. In: Journal of Biological Chemistry. 2019 ; Vol. 294, No. 7. pp. 2365-2374.
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Casein kinase II–mediated phosphorylation of lipin 1 phosphatidate phosphatase at Ser-285 and Ser-287 regulates its interaction with 14-3-3 protein. / Hennessy, Meagan; Granade, Mitchell E.; Hassaninasab, Azam; Wang, Dana; Kwiatek, Joanna M.; Han, Gil Soo; Harris, Thurl E.; Carman, George.

In: Journal of Biological Chemistry, Vol. 294, No. 7, 01.01.2019, p. 2365-2374.

Research output: Contribution to journalArticle

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T1 - Casein kinase II–mediated phosphorylation of lipin 1 phosphatidate phosphatase at Ser-285 and Ser-287 regulates its interaction with 14-3-3 protein

AU - Hennessy, Meagan

AU - Granade, Mitchell E.

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