title: Protein kinase C phosphorylation of PLCβ1 regulates its cellular localization.
abstract: Activation of phospholipase Cβ (PLCβ) by G proteins leads to a chain of events that result in an increase in intracellular calcium and activation of protein kinase C (PKC). It has been found that PKC phosphorylates PLCβ1 on S887 in vitro without affecting its enzymatic activity or its ability to be activated by Gα(q) proteins. To understand whether S887 phosphorylation affects the enzyme's activity in cells, we constructed two mutants that mimic the wild type and PKC-phosphorylated enzymes (S887A and S887D). We find that these constructs bind similarly to Gα(q) in vitro. When expressed in HEK293 cells, both mutants associate identically to Gα(q) in both the basal and stimulated states. Both mutants diffuse with similar rates and also interact identically with another known binding partner, translin-associated factor X (TRAX), which associates with PLCβ1 in the cytosol and nucleus. However, the two mutants localize differently in the cell. We find that S887A has a much higher nuclear localization than its S887D counterpart both in HEK293 cells and PC12 cells. Our studies suggest that PKC phosphorylation regulates the level of PLCβ1 cytosolic and nuclear activity by regulating its cellular compartmentalization.
authors.affiliation: Department of Physiology and Biophysics, Stony Brook University, Stony Brook, NY 11794-8661, USA.
authors.forename: Omozuanvbo
authors.initials: O
authors.lastname: Aisiku
authors.forename: Louisa
authors.initials: L
authors.lastname: Dowal
authors.forename: Suzanne
authors.initials: S
authors.lastname: Scarlata
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008958:Models, Molecular; D016716:PC12 Cells; D054799:Phospholipase C beta / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D017354:Point Mutation; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats
chemical_list: D004268:DNA-Binding Proteins; C498305:TSNAX protein, human; D011493:Protein Kinase C; D054799:Phospholipase C beta