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.

Human immunodeficiency virus type 1 capsid protein is a substrate of the retroviral proteinase while integrase is resistant toward proteolysis.

The capsid protein of human immunodeficiency virus type 1 was observed to undergo proteolytic cleavage in vitro when viral lysate was incubated in the presence of dithiothreitol at acidic pH. Purified HIV-1 capsid protein was also found to be a substrate of the viral proteinase in a pH-dependent manner; acidic pH (<7) was necessary for cleavage, and decreasing the pH toward 4 increased the degree of processing. Based on N-terminal sequencing of the cleavage products, the capsid protein was found to be cleaved at two sites, between residues 77 and 78 as well as between residues 189 and 190. Oligopeptides representing these cleavage sites were also cleaved at the expected peptide bonds. The presence of cyclophilin A decreased the degree of capsid protein processing. Unlike the capsid protein, integrase was found to be resistant toward proteolysis in good agreement with its presence in the preintegration complex.