Immunocytochemical localization of phospholipase C-gamma in rat embryo fibroblasts.

Rat embryo fibroblasts (REF52) exhibit a distinctive, transformation-sensitive distribution of alpha-protein kinase C (alpha-PKC). Receptor-mediated activation of phospholipase C (PLC)-gamma generates diacylglycerol, the major cellular activator of PKC. Immunofluorescence techniques were used to investigate the subcellular localization of two PLC isozymes (PLC-gamma and PLC-delta) in normal and simian virus 40-transformed REF52 cells to determine (i) if PLC colocalizes with alpha-PKC and (ii) if PLC isozyme distribution is sensitive to transformation. PLC-delta was not detected in either cell type. In REF52 cells, PLC-gamma was associated with the actin cytoskeleton and was evenly distributed along the length of the actin microfilaments. PLC-gamma was coincident with alpha-PKC at the points where the filaments are anchored to the membrane (i.e., the focal contacts). Cytoskeletal association of PLC-gamma was not transformation sensitive, although the actin cytoskeleton was more disordered in simian virus 40-transformed cells. In REF52 cells, platelet-derived growth factor induced tyrosine phosphorylation of both soluble and cytoskeletal PLC-gamma. Tyrosine phosphorylation of PLC-gamma did not seem to be a determinant of its subcellular localization, but there was a detectable increase in cytoskeleton-associated PLC-gamma in response to platelet-derived growth factor treatment.