The human papillomavirus type 16 E5 protein modulates phospholipase C-gamma-1 activity and phosphatidyl inositol turnover in mouse fibroblasts.
The human papillomavirus type 16 E5 (HPV16-E5) protein is a membrane protein that has been associated with malignant growth. The protein affects growth factor-mediated signal transduction in a ligand-dependent manner. We show now that E5 expression in A31 fibroblasts results in an increased level of diacylglycerol (DAG) and inositol phosphates. Immunoprecipitation of phospholipase C-gamma-1 (PLC-gamma-1) with specific antibodies and immunoblotting with anti-phosphotyrosine antibodies reveal a large increase in tyrosine phosphorylation of the enzyme in E5-expressing cells compared to control vector-transfected cells. This activation of tyrosine phosphorylation is growth factor independent. In addition, an enhanced formation of phosphatidic acid (PA) was observed in E5 cells. This increase did not result from activation of phospholipase D (PLD), although the enzyme was activatable by treatment with phorbol ester Thus, a phosphohydrolase-mediated DAG synthesis from PLD-produced PA can be excluded. The observed effects were not further enhanced by EGF showing that the presence of the growth factor is not necessary for maintaining permanent activation of PLC-gamma-1 in E5-expressing cells. The DAG- and inositol phosphate-mediated signal cascade within the cells is thus effectively uncoupled from external control via EGF and its receptor in the presence of E5 protein.
Crusius
K
K
Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Kaszkin
M
M
Kinzel
V
V
Alonso
A
A
D000818:Animals; D002460:Cell Line; D004075:Diglycerides / Q000378:metabolism; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005347:Fibroblasts / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D051379:Mice; D009856:Oncogene Proteins, Viral / Q000502:physiology*; D027383:Papillomaviridae / Q000502:physiology*; D010716:Phosphatidylinositols / Q000378:metabolism*; D051966:Phospholipase C gamma; D015398:Signal Transduction; D010738:Type C Phospholipases / Q000378:metabolism*; D000086082:Viroporin Proteins
D004075:Diglycerides; D004815:Epidermal Growth Factor; D007527:Isoenzymes; D009856:Oncogene Proteins, Viral; D010716:Phosphatidylinositols; D051966:Phospholipase C gamma; D010738:Type C Phospholipases; C054859:oncogene protein E5, Human papillomavirus type 16; D000086082:Viroporin Proteins