arkadia enhances nodaltgf-β signaling by coupling phospho-smad23 activity and turnoverarkadia增强nodaltgf-β信号耦合phospho-smad23活动和营业额.pdfVIP

PLoS BIOLOGY Arkadia Enhances Nodal/TGF-b Signaling by Coupling Phospho-Smad2/3 Activity and Turnover 1[ 1[ 1 1 1 Konstantinos J. Mavrakis , Rebecca L. Andrew , Kian Leong Lee , Chariklia Petropoulou , James E. Dixon , 1 2 1* Naveenan Navaratnam , Dominic P. Norris , Vasso Episkopou 1 Mammalian Neurogenesis, Medical Research Council, Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, London, United Kingdom, 2 Mammalian Genetics Unit, Medical Research Council, Harwell, United Kingdom Regulation of transforming growth factor-b (TGF-b) signaling is critical in vertebrate development, as several members of the TGF-b family have been shown to act as morphogens, controlling a variety of cell fate decisions depending on concentration. Little is known about the role of intracellular regulation of the TGF-b pathway in development. E3 ubiquitin ligases target specific protein substrates for proteasome-mediated degradation, and several are implicated in signaling. We have shown that Arkadia, a nuclear RING-domain E3 ubiquitin ligase, is essential for a subset of Nodal functions in the embryo, but the molecular mechanism of its action in embryonic cells had not been addressed. Here, we find that Arkadia facilitates Nodal signaling broadly in the embryo, and that it is indispensable for cell fates that depend on maximum signaling. Loss of Arkadia in embryonic cells causes nuclear accumulation of phospho-Smad2/3 (P-Smad2/3), the effectors of Nodal signaling; however, these must be repressed or hypoactive as the expression of their direct target genes is reduced or lost. Molecular and functional analysis shows that Arkadia interacts with and ubiquitinates P-Smad2/3 causing their degradation,