A 120 kDa nuclear phospholipase Cgamma1 protein fragment is stimulated in vivo by EGF signal phosphorylating nuclear membrane EGFR.
Intraperitoneal injection of epidermal growth factor (EGF) into mice resulted in the phosphorylation of liver nuclei phospholipase Cgamma1 (PLCgamma1) at the tyrosine, coincident with the time course of nuclear membrane epidermal growth factor receptor (EGFR) activation. The function of PLCgamma1 in mice liver nuclei was attributed to a 120 kDa protein fragment. This 120 kDa protein was immunoprecipitated with the isozyme specific PLCgamma1 antibody and was found to be sensitive to a PLCgamma1 specific blocking peptide. The 10-partial sequence analysis revealed that the 120 kDa protein contains the PELCQVSLSE sequence at its N-terminal end and the RTRVNGDNRL sequence at its C-terminal end, which reveals that this protein is a major fragment of PLCgamma1 devoid of an amino acid portion at the N-terminal end. The tyrosine-phosphorylated 120 kDa protein interacts with activated EGFR, binds phosphatidylinositol-3-OH-kinase enhancer (PIKE), enhances nuclear phosphatidylinositol-3-OH-kinase (PI[3]K) activity, and generates diacylglycerol (DAG) in response to the EGF signal to the nucleus in vivo. The immunoprecipitated 120 kDa protein fragment displayed phosphatidylinositol (PI) hydrolysis activity. These results establish the capacity of EGF-triggered nuclear signaling which is mediated by EGFR itself, located on the inner nuclear membrane. This is the first report identifying a 120 kDa PLCgamma1 fragment generated in vivo in the nucleus and capable of discharging the function of nuclear PLCgamma1.
Klein
Christian
C
Centre de Neurochimie du CNRS, Strasbourg, France.
Gensburger
Claire
C
Freyermuth
Solange
S
Nair
Bala C
BC
Labourdette
Gérard
G
Malviya
Anant N
AN
D000818:Animals; D002467:Cell Nucleus / Q000276:immunology / Q000378:metabolism; D004075:Diglycerides / Q000096:biosynthesis; D004789:Enzyme Activation / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology* / Q000502:physiology; D066246:ErbB Receptors / Q000378:metabolism*; D008099:Liver / Q000201:enzymology; D051379:Mice; D009685:Nuclear Envelope / Q000276:immunology / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology; D010716:Phosphatidylinositols / Q000378:metabolism; D051966:Phospholipase C gamma; D010766:Phosphorylation / Q000187:drug effects; D015398:Signal Transduction; D010738:Type C Phospholipases / Q000032:analysis / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism
D004075:Diglycerides; D010716:Phosphatidylinositols; D014443:Tyrosine; D004815:Epidermal Growth Factor; D019869:Phosphatidylinositol 3-Kinases; D066246:ErbB Receptors; D010738:Type C Phospholipases; D051966:Phospholipase C gamma