Ilphosphorylation. Rolipram ZK 62711 E13.5 mouse embryos was in the F Rbungsmuster both phosphorylated and phosphorylated Smad1 linker Smad1 tail / 5 mainly nuclear and showed a high level of Ma of colocalization. Phospho Smad1 linker and tail phosphorylated Smad1 / 5 were in the ventricular Ren recognized zone of the cerebral ventricles, hnen in the buds of Z, And in the medullary and spinal ganglia. Moderate levels were observed in the stomach wall, in the development of heart valves, the epithelial cells of the lungs and kidney tubules.

Phospho-Smad2 linker and tail phospho overlaps in the nuclei of the dorsal root ganglia stain, and only partially in male pattern germ cells localized CO, and ventricular areas in the brain and spinal cord Ren.
Smad2 phospho tail F Phospho staining with little or no linker was in the Z Hnen buds, mesenchymal cells around the airways, and a big heart valves e observed, the aortic wall, and the centers of Shortc Assurance of the vertebrae. In summary, the phosphorylation of Smad linker phosphorylation tail C is accompanied by a general feature of the BMP and TGF pathways. To determine the requirements for the ALP, we used mouse embryonic fibroblasts from wild-type embryos and embryos harvested homozygous for knock in Smad1 alleles with alanine mutations of the tail or C phosphorylation link. BMP did not induce ALP Smad1C intact despite the presence of this mutant in the linker sites, unlike UV irradiation of cells, the cytoplasmic Smad1 linker phosphorylation induced by JNK and p38 MAPK.
This indicates that the phosphorylation of Smad1 C-tail is not necessary for the phosphorylation of MAPK by binding antagonists are, but for in vivo phosphorylation by the binding of agonist-dependent Ngigen kinases. Smad ALP observed in all cell lines tested, was au It in cells lacking Smad4, phosphorylated pers Nlich general partner of Smad-activated receptor, which binds to the C tail bud formation and transcription complexes. Smad4 in SW480 defective c Lon line of human cancer and pancreatic cancer BxPC3 Online tail BMP-induced phosphorylation and nuclear accumulation of Smad1 / 5, but only minimal phosphorylation of Smad1 linker. Similar results were obtained with Smad3 in response to TGF. Restoring Smad4 expression rescued the F Ability of Smad1 and Smad3 on ALP to undergo. These results suggest that Smads undergo ALP after the incorporation phosphotail oriented transcription complexes with Smad4.
To determine whether there are Smad ALP in the regulatory regions of target genes, we performed Chromatinimmunpr Zipitation tests. BMP in the treated cells, but not controlled To an anti-Smad1 / 5 and an antique Body against phospho Ser206 of Smad1 folded regions of the DNA comprising the BMP-inhibitor react DNA binding and Smad7. In Similar way, in the TGF-treated cells, an antique Body against phosphorylated Smad3 linker and an anti-Smad2 / 3 folded DNA encoding the TGF-responsive element of the Smad7 gene. Treatment of cells with the inhibitor amanitin RNAP II had no effect on Smad1 ALP, indicating that this event is accompanied, but not a consequence of the active phosphorylated Smad1 is transcription.Linker recognized by Smurf1 and linkerphosphorylated Smad2 / 3 by NEDD4L, both go Ren to the family of HECT E3 ubiquitin ligases. The members of this family bind their substrates by WW dom