Siah proteins induce the epidermal growth factor-dependent degradation of phospholipase Cepsilon.
Phospholipase Cepsilon (PLCepsilon) is activated by various growth factors or G-protein-coupled receptor ligands via different activation mechanisms. The Ras association (RA) domain of PLCepsilon is known to be important for its ability to bind with Ras-family GTPase upon growth factor stimulation. In the present study, we identified Siah1 and Siah2 as novel binding partners of the PLCepsilon RA domain. Both Siah1 and Siah2 interacted with the RA2 domain of PLCepsilon, and the mutation of Lys-2186 of the PLCepsilon RA2 domain abolished this association. Moreover, Siah induced the ubiquitination and degradation of PLCepsilon upon epidermal growth factor (EGF) stimulation, and Siah proteins were phosphorylated on multiple tyrosine residues via an Src-dependent pathway upon EGF treatment. The Src inhibitor abolished the EGF-dependent ubiquitination of PLCepsilon, and the Siah1 phosphorylation-deficient mutant could not increase the EGF-dependent ubiquitination and degradation of PLCepsilon. The EGF-dependent degradation of PLCepsilon was blocked in mouse embryonic fibroblast (MEF) cells derived from Siah1a/Siah2 double knockout mice, and the extrinsic expression of wild-type Siah1 restored the degradation of PLCepsilon, whereas the phosphorylation-deficient mutant did not. Siah1 expression abolished PLCepsilon-dependent potentiation of EGF-dependent cell growth. In addition, the expression of wild-type Siah1 in Siah1a/Siah2-double knockout MEF cells inhibited EGF-dependent cell growth, and this inhibition was abolished by PLCepsilon knockdown. Our results suggest that the Siah-dependent degradation of PLCepsilon plays a role in the regulation of growth factor-dependent cell growth.
Yun
Sanguk
S
Department of Life Science, Division of Molecular and Life Science, Pohang University of Science and Technology, Pohang, Kyung-Buk 790-784, Republic of Korea.
Möller
Andreas
A
Chae
Suhn-Kee
SK
Hong
Won-Pyo
WP
Bae
Young Ju
YJ
Bowtell
David D L
DD
Ryu
Sung Ho
SH
Suh
Pann-Ghill
PG
D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D004622:Embryo, Mammalian; D004815:Epidermal Growth Factor / Q000502:physiology*; D005347:Fibroblasts / Q000502:physiology; D000882:Haplorhini; D006801:Humans; D007668:Kidney; D051379:Mice; D009687:Nuclear Proteins / Q000235:genetics / Q000502:physiology*; D054801:Phosphoinositide Phospholipase C / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000502:physiology*; D000098563:Seven in Absentia Proteins
D009687:Nuclear Proteins; D011994:Recombinant Proteins; D019000:Phosphotyrosine; D004815:Epidermal Growth Factor; D044767:Ubiquitin-Protein Ligases; D000098563:Seven in Absentia Proteins; D054801:Phosphoinositide Phospholipase C; C423320:phospholipase C epsilon