{PDOC51075} {PS51075; MH1} {PS51076; MH2} {BEGIN} ********************************************* * MAD homology domains MH1 and MH2 profiles * ********************************************* Smad proteins are signal transducers and transcriptional comodulators of the TGF-beta superfamily of ligands, which play a central role in regulating a broad range of cellular responses, including cell growth, differentiation, and specification of developmental fate, in diverse organisms from C. elegans to humans. Ligand binding to specific transmembrane receptor kinases induces receptor oligomerization and phosphorylation of the receptor specific Smad protein (R-Smad) in the cytoplasm. The R-Smad proteins regulate distinct signaling pathways. Smad1, 5 and 8 mediate the signals of bone morphogenetic proteins (BMPs), while Smad2 and 3 mediate the signals of activins and TGF- betas. Upon ligand stimulation, R-Smad proteins are phosphorylated at the conserved C-terminal tail sequence, SS*xS* (where S* denotes a site of phosphorylation). The phosphorylated states of R-Smad proteins form heteromeric complexes with Smad4 and are translocated into the nucleus. In the nucleus, the heteromeric complexes function as gene-specific transcription activators by binding to promoters and interacting with transcriptional coactivators. Smad6 and Smad7 are inhibitory Smad proteins that inhibit TGF- beta signaling by interfering with either receptor-mediated phosphorylation or hetero-oligomerization between Smad4 and R-Smad proteins. Smad proteins comprise two conserved MAD homology domains, one in the N-terminus (MH1) and one in the C-terminus (MH2), separated by a more variable, proline-rich linker region. The MH1 domain has a role in DNA binding and negatively regulates the functions of MH2 domain, whereas the MH2 domain is responsible for transactivation and mediates phosphorylation-triggered heteromeric assembly between Smad4 and R-Smad [1,2]. The MH1 domain adopts a compact globular fold, with four alpha helices, six short beta strands, and five loops (see ). The N-terminal half of the sequence consists of three alpha helices , and the C-terminal half contains all six beta strands, which form two small beta sheets and one beta hairpin. The fourth alpha helix is located in the hydrophobic core of the molecule, surrounded by the N-terminal three alpha helices on one side and by the two small beta sheets and the beta hairpin on the other side. These secondary structural elements are connected with five intervening surface loops. The MH1 domain employs a novel DNA-binding motif, a 11-residue beta- hairpin formed by strands B2 and B3, to contact DNA in the major groove. Two residues in the L3 loop and immediately preceding strand B2 also contribute significantly to DNA recognition. The beta hairpin appears to protrude outward from the globular MH1 core [1]. The MH2 domain is about ~230 amino acids in length and contains the SSxS motif. The MH2 domain is able to undergo phosphorylation-dependent homo- and heterotrimerization. The MH2 domain contains a central beta sandwich, with a conserved three helix bundle (H3, H4, and H5) on one end and a conserved loop- helix region (L1, L2, L3, and H1) on the other end (see ) [2]. The profiles we developed cover the entire MH1 and MH2 domains. -Sequences known to belong to this class detected by the profile: ALL. -Other sequence(s) detected in Swiss-Prot: NONE. -Sequences known to belong to this class detected by the profile: ALL. -Other sequence(s) detected in Swiss-Prot: NONE. -Note: It has been said that the MH1 domain is related to the CTF/NF-I DNA- binding domain (see ) [3,4]. -Last update: February 2005 / First entry. [ 1] Shi Y., Wang Y.-F., Jayaraman L., Yang H., Massague J., Pavletich N.P. "Crystal structure of a Smad MH1 domain bound to DNA: insights on DNA binding in TGF-beta signaling." Cell 94:585-594(1998). PubMed=9741623 [ 2] Wu J.-W., Hu M., Chai J., Seoane J., Huse M., Li C., Rigotti D.J., Kyin S., Muir T.W., Fairman R., Massague J., Shi Y. "Crystal structure of a phosphorylated Smad2. Recognition of phosphoserine by the MH2 domain and insights on Smad function in TGF-beta signaling." Mol. Cell 8:1277-1289(2001). PubMed=11779503 [ 3] Stefancsik R., Sarkar S. "Relationship between the DNA binding domains of SMAD and NFI/CTF transcription factors defines a new superfamily of genes." DNA Seq. 14:233-239(2003). PubMed=14631647 [ 4] Sadreyev R., Grishin N. "COMPASS: a tool for comparison of multiple protein alignments with assessment of statistical significance." J. Mol. Biol. 326:317-336(2003). PubMed=12547212 -------------------------------------------------------------------------------- PROSITE is copyrighted by the SIB Swiss Institute of Bioinformatics and distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives (CC BY-NC-ND 4.0) License, see https://prosite.expasy.org/prosite_license.html -------------------------------------------------------------------------------- {END}