The transforming growth factor beta (TGFbeta) family has critical roles in the regulation of fertility. In addition, the pathogenesis of some human cancers is attributed to misregulation of TGFbeta function and SMAD2 or SMAD4 mutations. There are limited mouse models for the BMP signaling SMADs (BR-SMADs) 1, 5, and 8 because of embryonic lethality and suspected genetic redundancy. Using tissue-specific ablation in mice, we deleted the BR-SMADs from somatic cells of ovaries and testes. Single conditional knockouts for Smad1 or Smad5 or mice homozygous null for Smad8 are viable and fertile. Female double Smad1 Smad5 and triple Smad1 Smad5 Smad8 conditional knockout mice become infertile and develop metastatic granulosa cell tumors. Male double Smad1 Smad5 conditional knockout mice are fertile but demonstrate metastatic testicular tumor development. Microarray analysis indicated significant alterations in expression of genes related to the TGFbeta pathway, as well as genes involved in infertility and

Protein and mRNA levels of SMAD3, but not of SMAD4 or SMAD7, were variably elevated in scleroderma fibroblasts compared with those from healthy controls. In sharp contrast to control fibroblasts, which displayed predominantly cytoplasmic localization of SMAD3/4 in the absence of exogenous TGFβ, in scleroderma fibroblasts SMAD3 and SMAD4 consistently showed elevated nuclear localization. Furthermore, phosphorylated SMAD2/3 levels were elevated and nuclear localization of phosphorylated SMAD2/3 was increased, suggesting activation of the SMAD pathway in scleroderma fibroblasts. Blockade of autocrine TGFβ signaling with antibodies or by expression of dominant-negative TGFβRII failed to normalize SMAD subcellular distribution, suggesting that elevated nuclear SMAD import was due to alterations downstream of the TGFβ receptors. The activity of a SMAD-responsive minimal promoter-reporter construct was enhanced in transiently transfected scleroderma fibroblasts. ...

By using tissue-specific knockout mouse model, we demonstrate for the first time that Smad2 is critical for VSMC differentiation from NCCs in vivo. The results are very specific because Smad2 deletion in NCCs only blocks VSMC marker gene expression in aortic arch arteries but not in the dorsal aorta. Although previous studies have tried to link Smad2 function with NCC differentiation to VSMCs or endothelial cells,22,44,45 the results are limited to the detection of Smad2 expression or use of in vitro systems. Our results represent the first direct evidence showing the critical role of Smad2 in VSMC differentiation from NCCs in vivo. Interestingly, Smad proteins seem to play a role in the progenitor-specific regulation of VSMC differentiation. Although both Smad2 and Smad3 mediate TGF-β function, Smad3 is important for the differentiation of mesenchymal-originated progenitors,21,41,42 whereas Smad2 plays a major role in VSMC differentiation from NCCs. Smad3 knockout mice are viable and show no ...

TY - JOUR. T1 - A novel Smad nuclear interacting protein, SNPIP1, suppresses p300- dependent TGF-β signal transduction. AU - Kim, Richard H.. AU - Wang, David. AU - Tsang, Michael. AU - Martin, Jennifer. AU - Huff, Carla. AU - De Caestecker, Mark P.. AU - Parks, W. Tony. AU - Meng, Xianwang. AU - Lechleider, Robert J.. AU - Wang, Tongwen. AU - Roberts, Anita B.. PY - 2000/7/1. Y1 - 2000/7/1. N2 - Members of the transforming growth factor-β superfamily play critical roles in controlling cell growth and differentiation. Effects of TGF-β family ligands are mediated by Smad proteins. To understand the mechanism of Smad function, we sought to identify novel interactors of Smads by use of a yeast two-hybrid system. A 396-amino acid nuclear protein termed SNIP1 was cloned and shown to harbor a nuclear localization signal (NLS) and a Forkhead-associated (FHA) domain. The carboxyl terminus of SNIP1 interacts with Smad1 and Smad2 in yeast two-hybrid as well as in mammalian overexpression systems. ...

The SMAD family of proteins in humans are homologs of Drosophila mothers against decapentaplegic and C. elegans Sma proteins. SMAD2 and SMAD3 are intracellular signaling proteins and transcriptional regulators that are activated by transforming growth factor beta (TGF-beta). Phosphorylation of SMAD2/3 is useful for understanding of various biological functions of TGF-beta. TGF-beta type I receptor and cyclin-dependent kinases phosphorylate both SMAD2 and SMAD3 in C-terminal and linker regions. TGF-beta signaling is mediated through these phosphorylation events. ...

The SMAD family of proteins in humans are homologs of Drosophila mothers against decapentaplegic and C. elegans Sma proteins. SMAD2 and SMAD3 are intracellular signaling proteins and transcriptional regulators that are activated by transforming growth factor beta (TGF-beta). Phosphorylation of SMAD2/3 is useful for understanding of various biological functions of TGF-beta. TGF-beta type I receptor and cyclin-dependent kinases phosphorylate both SMAD2 and SMAD3 in C-terminal and linker regions. TGF-beta signaling is mediated through these phosphorylation events. ...

Although hematopoiesis has been extensively studied, the molecular program of hematopoietic stem cell induction and self-renewal remains unclear. Recent work has revealed that the post-transcriptional processing of BMP Smads is involved in the maintenance of hematopoietic stem cell identity. Jiang et al. identified a C-terminal truncation in SMAD5, SMAD5β, which inactivates the protein. SMAD5β is expressed in both normal hematopoiesis and leukemogenesis (Jiang et al., 2000). Interestingly, the expression level of the SMAD5β isoform is higher in CD34+ hematopoietic stem cells than in terminally differentiated peripheral blood leukocytes, indicating that the alternative splicing of SMAD5 is differentially regulated during the maturation of hematopoietic cells. Our work shows that the splicing of smad1/5 is impaired by the knockdown of Ints5. Aberrant splicing of smad5 leads to the production of dominant-negative forms of Smad5, the overexpression of which phenocopies the hematopoiesis ...

The goal of this study is to define the function of Smurf2 in the regulation of TGF‐β signalling. Since the original discovery of Smurfs as ubiquitin ligases of Smads and TβRI, conflicting reports emerged in the literature as to whether Smurf2 promote Smad ubiquitination under physiological conditions. To resolve this issue, we generated mice carrying Smurf2‐deficient alleles, and have confirmed that Smurf2 is a physiological inhibitor of TGF‐β‐induced, Smad‐dependent transcriptional responses. However, instead of marking Smad3 for proteasomal degradation by poly‐ubiquitination, Smurf2 specifically promotes multiple mono‐ubiquitination of Smad3, which blocks formation of both homotrimeric Smad3 and heterotrimeric Smad3-Smad4 complexes.. Smurf1 and Smurf2 share a high degree of amino‐acid sequence homology. Mice with a targeted disruption of either the Smurf1 or Smurf2 allele are viable and survive to adulthood; however, disruption of both alleles simultaneously leads to ...

Smad2 is a crucial component of intracellular signaling by transforming growth factor-b (TGFb). Here we describe that Smad2 is glycosylated, which is a novel for Smad2 post-translational modification. We showed that the Smad2 glycosylation was inhibited upon treatment of cells with 17b-estradiol, and was enhanced in cells in a dense culture as compared to cells in a sparse culture. The Smad2 glycosylation was not dependent on the C-terminal phosphorylation of Smad2, and was not affected by TGFb1 treatment of the cells. Smad2 was glycosylated at multiple sites, and one of the predicted sites is Seine 110. Thus, Smad2 is glycosylated, and this post-translational modification was modulated by 17b-estradiol but not by TGFb1. ...

KIMURA, Edna T.; MATSUO, Sílvia E. and RICARTE-FILHO, Júlio Cézar. TGFb, activin and SMAD signalling in thyroid cancer. Arq Bras Endocrinol Metab [online]. 2007, vol.51, n.5, pp.683-689. ISSN 1677-9487. http://dx.doi.org/10.1590/S0004-27302007000500005.. TGFb and activin are members of the TGFb superfamily and play a wide role in development, proliferation and apoptosis. These growth factors exert their biological effects by binding to the type I and II membrane receptors to transduce their signalling through the nucleus by phosphorylation of R-SMADs (SMAD 2/3) and co-SMADs (Smad 4). The proper control of TGFb/activin pathway is negatively regulated by inhibitory SMAD (SMAD7) and by E3 ubiquitination enzymes (Smurfs). Physiologically, TGFb and activin act as potent growth inhibitors in thyroid follicular cell. Thus, alterations in the receptors and components of SMAD signalling pathway are associated with several types of tumors. Since TGFb and activin generate their intracellular signalling ...

SMADs function with other proteins to mediate transcriptional activation in response to transforming growth factor β and related proteins. SMADs are activated when phosphorylated by the TGF-β receptor, so one might have thought that the easy way to inactivate SMADs would be to dephosphorylate them. Although dephosphorylation might be a factor, new evidence points instead to a major role for ubiquitin-dependent degradation in termination of SMAD signaling. Lo and Massagué find that degradation of SMAD2 is increased in response to activation of the TGF-β receptor. Unlike regulation of some other transcription factors for which phosphorylation is a key tag that leads to ubiquitination and destruction, the essential event that leads to degradation of SMAD2 appears to be its translocation to the nucleus. The regulation of SMAD2 is also distinct from that recently described by Zhu et al. for SMAD1 and SMAD5, which appears to rely not on nuclear localization, but rather a cytoplasmic event that ...

Colorectal cancer (CRC) is one of the most common causes of cancer related deaths in western countries. CRC are commonly divided in cancers showing microsatellite stability (MSS) or microsatellite instability (MSI). A more novel classification is dependent on promoter hypermethylation of CpG islands (the CpG island methylator phenotype, CIMP), where cancers show high, low or negative methylation status. SMAD4, located on chromosome 18q, has been thoroughly investigated during the last years. Loss of SMAD4 expression has been reported to correlate with poor CRC patient prognosis. In this study we analyze the impact of SMAD4 expression on prognosis in relation to MSI screening status and CIMP status. 479 paraffin-embedded specimens of CRC were examined for nuclear SMAD4 expression using immunohistochemistry. The tumors were scored loss (-), moderate (+) and high (++) expressing tumors. Loss of SMAD4 correlated significantly with decreased survival in all colon cancer patients. High SMAD4 ...

Background: Smad1 signaling is essential for cardiovascular development. It was previously reported that cardiac specific Smad1 overexpression results in a myocardio-protective effect in myocardial ischemia/reperfusion (I/R). However, it remains unclear if Smad1 signaling is required for an adult heart.. Methods and results: To investigate a role of Smad1 signaling in cardiac pathophysiology, we generated a cardiomyocyte-restricted Smad1 knockout line (CS1K) using Cre-loxP technology. A previously published mouse line with cardiomyocyte-restricted Cre overexpression (α-MyHC-Cre) was used for the crossing and as control. Smad1 was largely eliminated in cardiomyocytes isolated from CS1K mice. The CS1K mice were overtly normal during the first 5- 6 months after birth. No pathophysiological abnormality could be detected during this period of time. However, the CS1K mice manifested cardiac hypertrophy at around 6 months and heart failure at later time points. Heart weight to body weight ratio of ...

Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD1 is a receptor-regulated SMAD (R-SMAD) (By similarity). May play a role in the initiation and maintenance of spermatogenesis. SMAD1/OAZ1/PSMB4 complex mediates the degradation of the CREBBP/EP300 repressor SNIP1 (By similarity). May act synergistically with SMAD4 and YY1 in bone morphogenetic protein (BMP)-mediated cardiac-specific gene expression (PubMed:15329343).

Transforming growth factor-beta (TGF-beta) signalling is induced in liver as a consequence of damage and contributes to wound healing with transient activation, whereas it mediates fibrogenesis with long-term up-regulation in chronic disease. Smad-dependent TGF-beta effects are blunted by antagonistic Smad7, which is transcriptionally activated as an immediate early response upon initiation of TGF-beta signalling in most cell types, thereby providing negative feedback regulation. Smad7 can be induced by other cytokines, e.g. IFN-gamma, leading to a crosstalk of these signalling pathways. Here we report on a novel mouse strain, denoted S7DeltaE1, with a deletion of exon I from the endogenous smad7 gene. The mice were viable and exhibited normal adult liver architecture. To obtain insight into Smad7-depend-ent protective effects, chronic liver damage was induced in mice by carbon tetrachloride (CCI4) administration. Subsequent treatment, elevated serum liver enzymes indicated enhanced liver damage ...

View Amhr2/Amhr2 Smad1/Smad1 Smad5/Smad5 involves: 129P2/OlaHsd * 129S/SvEv * C57BL/6J: phenotypes, images, diseases, and references.

View Amhr2/Amhr2 Smad1/Smad1 Smad5/Smad5 involves: 129P2/OlaHsd * 129S/SvEv * C57BL/6J: phenotypes, images, diseases, and references.

The TGFβ signaling pathway plays essential roles during embryogenesis (Massagué and Chen, 2000; Hill, 2001; Muñoz-Sanjuán and Hemmati-Brivanlou, 2001; Whitman, 2001). Members of the TGFβ family exert their biological functions by binding to two types of transmembrane receptors, type I and type II, that encode a serine/threonine kinase in the intracellular domain. Upon ligand binding, the type II receptor phosphorylates the type I receptor, which subsequently activates members of the Smad family of intracellular signal transducers in the cytosol. Ligands of the TGFβ family are subdivided into a few groups and each group activates different sets of the receptor-regulated Smad (R-Smad). Activin/TGFβ signals through Smad2 and Smad3, whereas bone morphogenetic proteins (BMPs) use Smad1, Smad5 and Smad8. Smad4, also known as the common-partner Smad (Co-Smad), forms a heteromeric complex with the R-Smads in response to ligand stimulation and is commonly used by both activin/TGFβ and BMPs. Upon ...

Activation of transforming growth factor-β (TGF-β) and activin receptors leads to phosphorylation of Sma- and Mad-related protein 2 (Smad2) and Smad3, which function as transcription factors to regulate gene expression. Smad7 is a regulatory protein which is able to inhibit TGF-β and activin signalling in a negative-feedback loop, mediated by a direct regulation by Smad3 and Smad4 via a Smad-binding element (SBE) in the Smad7 promoter. Interestingly, we found that the Smad7 promoter was also regulated by nuclear factor ĸB (NF-ĸB), a transcription factor which plays an important role in inflammation and the immune response. Expression of NF-ĸB p65 subunit was able to inhibit the Smad7 promoter activity, and this inhibition could be reversed by co-expression of IĸB, an inhibitor of NF-ĸB. In addition, the inhibitory activity of p65 was observed in a minimal promoter that contained only the Smad7 SBE and a TATA box, without any consensus NF-ĸB binding site. This inhibitory effect appeared ...

Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD2/SMAD4 complex, activates transcription. May act as a tumor suppressor in colorectal carcinoma. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator (By similarity).

The transforming growth factor-beta (TGF-beta) family members, which include TGF-betas, activins and bone morphogenetic proteins (BMPs), are structurally related secreted cytokines found in species ranging from worms and insects to mammals. A wide spectrum of cellular functions such as proliferation, apoptosis, differentiation and migration are regulated by TGF-beta family members. TGF-beta family member binds to the Type II receptor and recruits Type I, whereby Type II receptor phosphorylates and activates Type I. The Type I receptor, in turn, phosphorylates receptor-activated Smads ( R-Smads: Smad1, Smad2, Smad3, Smad5, and Smad8). Once phosphorylated, R-Smads associate with the co-mediator Smad, Smad4, and the heteromeric complex then translocates into the nucleus. In the nucleus, Smad complexes activate specific genes through cooperative interactions with other DNA-binding and coactivator (or co-repressor) proteins ...

The transforming growth factor-beta (TGF-beta) family members, which include TGF-betas, activins and bone morphogenetic proteins (BMPs), are structurally related secreted cytokines found in species ranging from worms and insects to mammals. A wide spectrum of cellular functions such as proliferation, apoptosis, differentiation and migration are regulated by TGF-beta family members. TGF-beta family member binds to the Type II receptor and recruits Type I, whereby Type II receptor phosphorylates and activates Type I. The Type I receptor, in turn, phosphorylates receptor-activated Smads ( R-Smads: Smad1, Smad2, Smad3, Smad5, and Smad8). Once phosphorylated, R-Smads associate with the co-mediator Smad, Smad4, and the heteromeric complex then translocates into the nucleus. In the nucleus, Smad complexes activate specific genes through cooperative interactions with other DNA-binding and coactivator (or co-repressor) proteins ...

993 Background: SMAD4 gene is a tumor suppressor that is critical for transmitting signals from transforming growth factor-beta and is associated with cross-talk of signal pathways. We investigated the expression of SMAD4 to evaluate the role of SMAD4 in the development and progression and to assess the clinical values after surgery in colorectal adenocarcinomas. Methods: Immunohistochemistry for SMAD4 was performed on the tissue microarray of 429 colorectal adenocarcinomas. Results: Two hundred three of 429 cases (47.3%) were positive and 226 cases (52.7%) were negative for SMAD4 expression. 2/10 (20.0%), 15/39 (38.5%), 68/114 (47.2%), 111/190 (58.4%), 30/46 (65.2%) cases were negative for SMAD4 expression in AJCC stage 0, I, II, III, IV, respectively. 138/226 (61.1%) of SMAD4 negative cases and 90/203 (44.3%) of SMAD4 positive cases had lymphatic metastasis (p=0.001). 148/226 (65.5%) of SMAD4 negative cases and 91/203 (44.8%) of SMAD4 positive cases had lymphovascular invasion (p,0.0001). T ...

The master cytokine TGF-β mediates tissue fibrosis associated with inflammation and tissue injury. TGF-β induces fibroblast activation and differentiation into myofibroblasts that secrete extracellular matrix proteins. Canonical TGF-β signaling mobilizes Smad2 and Smad3 transcription factors that control fibrosis by promoting gene expression. However, the importance of TGF-β-Smad2/3 signaling in fibroblast-mediated cardiac fibrosis has not been directly evaluated in vivo. Here, we examined pressure overload-induced cardiac fibrosis in fibroblast- and myofibroblast-specific inducible Cre-expressing mouse lines with selective deletion of the TGF-β receptors Tgfbr1/2, Smad2, or Smad3. Fibroblast-specific deletion of Tgfbr1/2 or Smad3, but not Smad2, markedly reduced the pressure overload-induced fibrotic response as well as fibrosis mediated by a heart-specific, latency-resistant TGF-β mutant transgene. Interestingly, cardiac fibroblast-specific deletion of Tgfbr1/2, but not Smad2/3, ...

4213 Transforming growth factor-? (TGF-?) is a multifunctional cytokine which signals to the nucleus through cell surface transmembrane serine/threonine kinase receptors and cytoplasmic effectors, including Smad2, Smad3 and Smad4 proteins. The role of TGF-? in carcinogenesis is complex with tumor suppressor activities in early stages of disease and pro-oncogenic activities in advanced stages of invasive, metastatic disease. We recently identified a novel modulator of this pathway, TLP (TRAP-1-like protein), which associates with TGF-? receptors constitutively and with the common-mediator Smad4 upon ligand binding. We have shown that TLP regulates formation of Smad3/4 complexes and Smad3-dependent transcriptional responses. Since no inactivating mutations in Smad3 have been observed in human tumors, we hypothesized that proteins that regulate Smad3 function, such as TLP, rather than Smad3 itself might be direct targets of oncogenic change. To assess whether the expression of TLP might be ...

p,Specific chromatin marks keep master regulators of differentiation silent yet poised for activation by extracellular signals. We report that nodal TGF-β signals use the poised histone mark H3K9me3 to trigger differentiation of mammalian embryonic stem cells. Nodal receptors induce the formation of companion Smad4-Smad2/3 and TRIM33-Smad2/3 complexes. The PHD-Bromo cassette of TRIM33 facilitates binding of TRIM33-Smad2/3 to H3K9me3 and H3K18ac on the promoters of mesendoderm regulators Gsc and Mixl1. The crystal structure of this cassette, bound to histone H3 peptides, illustrates that PHD recognizes K9me3, and Bromo binds an adjacent K18ac. The interaction between TRIM33-Smad2/3 and H3K9me3 displaces the chromatin-compacting factor HP1γ, making nodal response elements accessible to Smad4-Smad2/3 for Pol II recruitment. In turn, Smad4 increases K18 acetylation to augment TRIM33-Smad2/3 binding. Thus, nodal effectors use the H3K9me3 mark as a platform to switch master regulators of stem cell ...

The master cytokine TGF-β mediates tissue fibrosis associated with inflammation and tissue injury. TGF-β induces fibroblast activation and differentiation into myofibroblasts that secrete extracellular matrix proteins. Canonical TGF-β signaling mobilizes Smad2 and Smad3 transcription factors that control fibrosis by promoting gene expression. However, the importance of TGF-β-Smad2/3 signaling in fibroblast-mediated cardiac fibrosis has not been directly evaluated in vivo. Here, we examined pressure overload-induced cardiac fibrosis in fibroblast- and myofibroblast-specific inducible Cre-expressing mouse lines with selective deletion of the TGF-β receptors Tgfbr1/2, Smad2, or Smad3. Fibroblast-specific deletion of Tgfbr1/2 or Smad3, but not Smad2, markedly reduced the pressure overload-induced fibrotic response as well as fibrosis mediated by a heart-specific, latency-resistant TGF-β mutant transgene. Interestingly, cardiac fibroblast-specific deletion of Tgfbr1/2, but not Smad2/3, ...

The master cytokine TGF-β mediates tissue fibrosis associated with inflammation and tissue injury. TGF-β induces fibroblast activation and differentiation into myofibroblasts that secrete extracellular matrix proteins. Canonical TGF-β signaling mobilizes Smad2 and Smad3 transcription factors that control fibrosis by promoting gene expression. However, the importance of TGF-β-Smad2/3 signaling in fibroblast-mediated cardiac fibrosis has not been directly evaluated in vivo. Here, we examined pressure overload-induced cardiac fibrosis in fibroblast- and myofibroblast-specific inducible Cre-expressing mouse lines with selective deletion of the TGF-β receptors Tgfbr1/2, Smad2, or Smad3. Fibroblast-specific deletion of Tgfbr1/2 or Smad3, but not Smad2, markedly reduced the pressure overload-induced fibrotic response as well as fibrosis mediated by a heart-specific, latency-resistant TGF-β mutant transgene. Interestingly, cardiac fibroblast-specific deletion of Tgfbr1/2, but not Smad2/3, ...

Lopez-Coviella, I., Mellott, T. M., Kovacheva, V. P., Berse, B., Slack, B. E., Zemelko, V., ... Blusztajn, J. K. (2006). Developmental pattern of expression of BMP receptors and Smads and activation of Smad1 and Smad5 by BMP9 in mouse basal forebrain. Brain Research, 1088(1), 49 - 56 ...

The TGF-β signaling pathway fulfills important roles in the normal and pathological function of many cells and tissues (1, 2). Our study describes an in vivo reporter mouse for this pathway and specifically for activators of Smad2/3. We demonstrate that this pathway has a high basal activity in the brain, intestine, heart, and skin and that it is rapidly activated after brain injury and inflammation.. Smad2 and Smad3 mediate transcriptional responses for TGF-β, nodal, activin, and some growth and differentiation factors (2, 4), and the SBE-luc reporter gene used here seems to respond to all of these tested in primary astrocytes derived from the reporter mice (Fig. 2⇑B). Activation of the reporter gene in vivo at basal state or after injury may therefore reflect signaling by either one or combinations of these growth factors and future studies using targeted inactivation of specific receptors or ligands will have to dissect these responses. At the transcriptional level, the reporter gene may ...

PDLIM5 activates SMAD3 via binding. STUB1 inhibits SMAD3 via ubiquitination. : PDLIM5 inhibits STUB1-mediated degradation of SMAD3 and promotes the migration and invasion of lung cancer cells.

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Here, we present several novel important findings as follows: First, activation of CD137 signaling induces NFATc1 expression and promotes angiogenesis in vivo, in vitro, and ex vivo. Second, CD137 signaling regulates phosphorylation of Smad1/5 and NFATc1 expression in ECs. Silencing of Smad1/5 attenuates the expression of NFATc1 and angiogenesis in vitro induced by CD137 signaling. Suppressing NFATc1 gene could inhibit angiogenesis induced by CD137 signaling in vitro. Therefore, CD137 signaling regulates angiogenesis by modulating Smad1/5‐NFATc1 signaling.. Atherosclerosis, a disease of chronic inflammation, is the primary cause of heart disease and stroke. As an important member of the TNFR superfamily, CD137‐CD137L interaction promotes inflammation. Evidence suggests that activation of CD137‐CD137L signaling may be a significant triggering event for the process of forming atherosclerotic plaque, and contributes to the plaque vulnerability.4 However, whether activation of CD137 has the ...

Rabbit anti SMAD3 antibody recognizes SMAD3, also known as mothers against decapentaplegic homolog 3. SMAD3 is a transcription factor bel

S with PBMCs could promote the expression of Smad2 and Smad3. This suggests that a Smaddependent mechanism might be existed in gastric tumor microenvironment.

Because the Smad-dependent pathway, the canonical TGF-β signaling mechanism, is activated under several pathological conditions in the heart,13 it was postulated that the Smad-dependent pathway mediates detrimental functions of TGF-β1. However, recent studies showed that this hypothesis may not necessarily be true. Wang et al reported a surprising observation that cardiac-specific Smad4 knockout mice resulted in cardiac hypertrophy and heart failure.14 Because Smad4 is considered the common transcriptional mediator of the Smad-dependent pathway, the observation by Wang et al suggests that the Smad-dependent pathway rather inhibits cardiac hypertrophy, apoptosis, and fibrosis, thereby acting in a protective manner. Although this observation does not exclude the participation of active Smads in hypertrophy and failure, the disadvantage caused by the lack of Smad4 overwhelms the few, if any, advantages to abolishing active Smad signaling in the heart. Xu et al found that Smad2 and Smad3 are ...

TY - JOUR. T1 - Histone 3 lysine 9 (H3K9) methyltransferase recruitment to the interleukin-2 (IL-2) promoter is a mechanism of suppression of IL-2 transcription by the transforming growth factor-β-smad pathway. AU - Wakabayashi, Yu. AU - Tamiya, Taiga. AU - Takada, Ichiro. AU - Fukaya, Tomohiro. AU - Sugiyama, Yuki. AU - Inoue, Naoko. AU - Kimura, Akihiro. AU - Morita, Rimpei. AU - Kashiwagi, Ikko. AU - Takimoto, Tomohito. AU - Nomura, Masatoshi. AU - Yoshimura, Akihiko. PY - 2011/10/14. Y1 - 2011/10/14. N2 - Suppression of IL-2βproduction from T cells is an important process for the immune regulation by TGF-β. However, the mechanism by which this suppression occurs remains to be established. Here, we demonstrate that Smad2 and Smad3, two major TGF-β-downstream transcription factors, are redundantly essential for TGF-β-mediated suppression of IL-2 production in CD4 + T cells using Smad2- and Smad3-deficient T cells. Both Smad2 and Smad3 were recruited into the proximal region of the IL-2 ...

The transforming growth factor beta (TGFB) signaling pathway is involved in many cellular processes in both the adult organism and the developing embryo including cell growth, cell differentiation, apoptosis, cellular homeostasis and other cellular functions. In spite of the wide range of cellular processes that the TGFβ signaling pathway regulates, the process is relatively simple. TGFβ superfamily ligands bind to a type II receptor, which recruits and phosphorylates a type I receptor. The type I receptor then phosphorylates receptor-regulated SMADs (R-SMADs) which can now bind the coSMAD SMAD4. R-SMAD/coSMAD complexes accumulate in the nucleus where they act as transcription factors and participate in the regulation of target gene expression ...

The Smad family of proteins, which are frequently targeted by tumorigenic mutations in cancer, mediate TGF-beta signaling from cell membrane to nucleus. The crystal structure of a Smad3 MH1 domain bound to an optimal DNA sequence determined at 2.8 A resolution reveals a novel DNA-binding motif. In the crystals, base-specific DNA recognition is provided exclusively by a conserved 11-residue beta hairpin that is embedded in the major groove of DNA. A surface loop region, to which tumorigenic mutations map, has been identified as a functional surface important for Smad activity. This structure establishes a framework for understanding how Smad proteins may act in concert with other transcription factors in the regulation of TGF-beta-responsive genes. Crystal structure of a Smad MH1 domain bound to DNA: insights on DNA binding in TGF-beta signaling.,Shi Y, Wang YF, Jayaraman L, Yang H, Massague J, Pavletich NP Cell. 1998 Sep 4;94(5):585-94. PMID:9741623[14] From MEDLINE®/PubMed®, a database of the ...

Rabbit polyclonal antibody raised against synthetic peptide of SMAD1. A synthetic peptide corresponding to N-terminus of human SMAD1. (PAB5579) - Products - Abnova

Fibrosis in inflammatory bowel disease is believed to be due to an excess deposition of extracellular matrix (ECM) by myofibroblasts and an impairment of the matrix degrading metalloproteinases (MMPs). Transforming growth factor-β (TGFβ) stimulates ECM synthesis and the level of MMPs are reduced by tissue inhibitors of metalloproteinase (TIMPs). This study examined uninflamed mucosa from 25 patients with Crohns disease (CD) with strictures and compared this with tissue from 18 patients with CD without stricturing. TGFβ was over expressed in myofibroblasts from mucosa overlying strictures (see fig). Binding of TGF activates the Smad proteins, phosphorylating Smad 2 and 3 whose action on gene transcription is inhibited by Smad 6 and 7. The authors showed an increase in phosphorylated Smad 2 and 3 from mucosa above the stricture and a decrease in the inhibitory Smad 6 and 7. There was also a reduction in mucosal MMP-12 and an increase in the inhibitor TIMP-1. Isolated myofibroblasts from ...

BMP receptors determine the intensity of BMP signals via Smad1 C-terminal phosphorylations. Here we show that a finely controlled cell biological pathway terminates this activity. The duration of the activated pSmad1(Cter) signal was regulated by sequential Smad1 linker region phosphorylations at co …

Background-Vascular aneurysm is an abnormal local dilatation of an artery that can lead to vessel rupture and sudden death. The only treatment involves surgical or endovascular repair or exclusion. There is currently no approved medical therapy for this condition. Recent data established a strong association between genetic variants in the 9p21 chromosomal region in humans and the presence of cardiovascular diseases, including aneurysms. However, the mechanisms linking this 9p21 DNA variant to cardiovascular risk are still unknown. Methods and Results-Here we show that deletion of the orthologous 70-kb non-coding interval on mouse chromosome 4 (chr4Δ70kb/Δ70kb mice) is associated with reduced aortic expression of cyclin-dependent kinase (CDK) inhibitors p19Arf and p15Inkb. Vascular smooth muscle cells from chr4Δ70kb/Δ70kb mice show reduced transforming growth factor (TGF)-β-dependent canonical Smad2 signaling but increased CDK-dependent Smad2 phosphorylation at linker sites, a phenotype ...

TGFβ1 enhanced Smad3 recruitment to the TGFBIp and ECM-associated gene promoters. Smad3 recruitment at the indicated gene promoters in wild-type and GCD2 homoz

Mouse monoclonal antibody raised against a full length recombinant SMAD1. SMAD1 (AAH01878.1, 1 a.a. ~ 465 a.a) full-length recombinant protein with GST tag. MW of the GST tag alone is 26 KDa. (H00004086-M04) - Products - Abnova

Rabbit polyclonal Smad2 + Smad3 antibody validated for WB, ELISA and tested in Human and Rat. Referenced in 4 publications. Immunogen corresponding to…

マウス・モノクローナル抗体 ab75273 交差種: Rat,Hu 適用: WB,ELISA,Flow Cyt,ChIP…SMAD1+SMAD5抗体一覧…画像、プロトコール、文献などWeb上の情報が満載のアブカムの Antibody…

Right here we also display that, as predicted, AB215 doesnt signal through SMAD2 3 and, consequently, does not signal in an Activin A like manner in HEK293T cells. We more examined the signaling properties of AB215 in human MCF7 breast cancer cells and discovered that, similar to what was observed in C2C12 cells, AB215 generates prolonged and enhanced Inhibitors,Modulators,Libraries SMAD1 five 8 phosphorylation when in contrast to that induced by BMP2. The amount of BMP2 induced SMAD1 5 eight phosphorylation in MCF7 cells peaks after 60 minutes then decreases to basal amounts right after 3 hours. By contrast, treatment of these cells with AB215 final results in maximal SMAD1 five 8 phosphorylation thirty min following stimulation and sustained immediately after six hrs.. We also made use of a reporter construct consisting from the phospho SMAD1 five eight responsive ID1 promoter upstream of the luciferase gene to compare the results of BMP2 and AB215 remedy over the human breast can cer cell ...

In this study, we demonstrated for the first time that Smad2 and Smad3 are essential for TGF-β-mediated regulation of T cell immunity. Smad2-cKO mice exhibited some immunologic disturbance, but not at lethal levels. When certain inflammatory events occurred, such as dextran sodium sulfate-induced colitis, Smad2-cKO mice failed to maintain the immunologic homeostasis. Furthermore, we showed that Smad2 and Smad3 have functional redundancy, sharing common roles in T cells. One Smad partially compensated for the deficiency of another in particularly important functions, such as Foxp3 induction or maintenance or Th1 cell suppression. Consistent with this idea, Smad2-three-dimensional-KO mice because of severe inflammation, as did TGF-β1 KO mice. We demonstrated in this study that TGF-β-mediated induction of Foxp3 is also dependent on both Smad2 and Smad3 signaling. Furthermore, we demonstrated that both Smad2 and Smad3 play an important role in the maintenance of Foxp3 expression in nTregs. Like ...

The expression and localisation of downstream signalling molecules of transforming growth factor beta superfamily, Smad2 and Smad4 proteins, was investigated in immature and mature dog testis. Cellular localisation of Smad2 and Smad4 proteins was examined using immunohistochemistry. Quantitative analysis of immunostaining was determined by the image analysis system. The specificity of the antibodies was examined using Western blotting assay. Smad2 and Smad4 were widely expressed in the testes, mainly immunolocalised in the cytoplasm of gonocytes, Leydig cells and Sertoli cells of immature testes, and Leydig cells and Sertoli cells of mature testes. At the same time, the expression levels for both proteins were different between immature and mature age groups: the expression of Smad2 and Smad4 proteins in the Sertoli cells of mature testis was higher than in the immature group (P | 0.05), but the expression of these two proteins in Leydig cells of mature testis was weaker than that seen in immature stage

TY - JOUR. T1 - Gastric tumor development in Smad3-deficient mice initiates from forestomach/glandular transition zone along the lesser curvature. AU - Nam, Ki Taek. AU - ONeal, Ryan. AU - Lee, Yeo Song. AU - Lee, Yong Chan. AU - Coffey, Robert J.. AU - Goldenring, James R.. N1 - Copyright: Copyright 2012 Elsevier B.V., All rights reserved.. PY - 2012/6. Y1 - 2012/6. N2 - SMAD proteins are downstream effectors of the TGF-Β signaling pathway. Smad3-null mice develop colorectal cancer by 6 months of age. In this study, we have examined whether the loss of Smad3 promotes gastric neoplasia in mice. The stomachs of Smad3 -/-mice were compared with age-matched Smad3 heterozygous and wild-type mice. E-cadherin, Ki-67, phosphoSTAT3, and TFF2/SP expression was analyzed by immunohistochemisty. The short hairpin RNA (ShRNA)-mediated knockdown of Smad3 in AGS and MKN28 cells was also performed. In addition, we examined alterations in DCLK1-expressing cells. Smad3/mouse stomachs at 6 months of age revealed ...

N-terminal Mad Homology 1 (MH1) domain. The MH1 is a small DNA-binding domain present in SMAD (small mothers against decapentaplegic) family of proteins, which are signal transducers and transcriptional modulators that mediate multiple signaling pathways. MH1 binds to the DNA major groove in an unusual manner via a beta hairpin structure. It negatively regulates the functions of the MH2 domain, the C-terminal domain of SMAD. Receptor-regulated SMAD proteins (R-SMADs, including SMAD1, SMAD2, SMAD3, SMAD5, and SMAD9) are activated by phosphorylation by transforming growth factor (TGF)-beta type I receptors. The active R-SMAD associates with a common mediator SMAD (Co-SMAD or SMAD4) and other cofactors, which together translocate to the nucleus to regulate gene expression. The inhibitory or antagonistic SMADs (I-SMADs, including SMAD6 and SMAD7) negatively regulate TGF-beta signaling by competing with R-SMADs for type I receptor or Co-SMADs. MH1 domains of R-SMAD and SMAD4 contain a nuclear ...

To better understand the roles of TGF-beta in bone metabolism, we investigated osteoclast survival in response TGF-beta and found that TGF-beta inhibited apoptosis. We examined the receptors involved in promotion of osteoclast survival and found that the canonical TGF-beta receptor complex is involved in the survival response. The upstream MEK kinase TAK1 was rapidly activated following TGF-beta treatment. Since osteoclast survival involves MEK, AKT, and NFkappaB activation, we examined TGF-beta effects on activation of these pathways and observed rapid phosphorylation of MEK, AKT, IKK, IkappaB, and NFkappaB. The timing of activation coincided with SMAD activation and dominant negative SMAD expression did not inhibit NFkappaB activation, indicating that kinase pathway activation is independent of SMAD signaling. Inhibition of TAK1, MEK, AKT, NIK, IKK, or NFkappaB repressed TGF-beta-mediated osteoclast survival. Adenoviral-mediated TAK1 or MEK inhibition eliminated TGF-beta-mediated kinase pathway

Smad7 is an intracellular antagonist of transforming growth factor-β signalling pathways and modulates muscle growth in vivo. Loss of Smad7 results in decreased muscle mass, reduced force generation, fibre type switching from glycolytic towards oxidative type and delayed recovery from injury. Upregulated Smad2/3 signalling in Smad7(-/-) muscle results in reduced myoblast proliferation and differentiation. Smad7 is an important regulator of muscle growth and may be a potential intracellular therapeutic target for muscle disorders.The transforming growth factor-β (TGF-β) family of growth factors plays an essential role in mediating cellular growth and differentiation. Myostatin is a muscle-specific member of the TGF-β superfamily and a negative regulator of muscle growth. Myostatin inhibitors are currently being pursued as therapeutic options for muscle disorders. Smad7 inhibits intracellular myostatin signalling via Smad2/3, and thus presents a means of regulating myostatin and potentiating ...

Background: Small mothers against decapentaplegic (SMAD)4 and SMAD7 are key regulatory components in the immunosuppressive transforming growth factor- (TGF-) β signaling pathway, which is defective in inflammatory bowel disease (IBD). SMAD4 may play an important role in the pathogenesis of IBD as indicated in experimental models of colitis.. Aims: To examine the ileal expression levels of SMAD4 and to correlate these with CD disease activity.. Methods: The material comprised 29 CD patients (13 with active disease, 16 in remission) and 9 asymptomatic patients referred for ileocolonoscopy as part of an adenoma surveillance program serving as controls. Patients were examined with ileocolonoscopy. Corresponding ileal biopsies were obtained for histological analysis and assessment of SMAD4 and SMAD7 protein expression by immunohistochemistry (IHC).. Results: The protein expression of SMAD4 was significantly downregulated in ileal tissue sections from CD patients as compared to healthy controls (p , ...

SMAD4, also called SMAD family member 4, Mothers against decapentaplegic homolog 4, or DPC4 (Deleted in Pancreatic Cancer-4) is a highly-conserved protein present in all metazoans. It belongs to the SMAD family of transcription factor proteins, which act as mediators of TGF-β signal transduction. The TGFβ family of cytokines regulates critical processes during the lifecycle of metazoans, with important roles during embryo development, tissue homeostasis, regeneration, and immune regulation. SMAD 4 belongs to the co-SMAD group, the second class of the SMAD family. SMAD4 is the only known co-SMAD in most metazoans. It also belongs to the Darwin family of proteins that modulate members of the TGFβ protein superfamily, a family of proteins that all play a role in the regulation of cellular responses. Mammalian SMAD4 is a homolog of the Drosophila protein Mothers against decapentaplegic named Medea. SMAD4 interacts with R-Smads, such as SMAD2, SMAD3, SMAD1, SMAD5 and SMAD8 (also called SMAD9) to ...

TY - JOUR. T1 - Essential role for Smad3 in angiotensin II-induced tubular epithelial-mesenchymal transition. AU - Yang, Fuye. AU - Huang, Xiao Ru. AU - Chung, Arthur C K. AU - Hou, Chun Cheng. AU - Lai, Kar Neng. AU - Lan, Hui Yao. PY - 2010/8. Y1 - 2010/8. N2 - Angiotensin II (Ang II) is a key mediator of chronic kidney disease, in which epithelial-mesenchymal transition (EMT) is a critical process mediated by the TGFβ/Smad signalling pathway. The present study examined the specific role of Smads in Ang II-induced EMT in vitro and in vivo. We found that Ang II signalled through the receptor of AT1, not AT2, to activate Smad2/3 and induce EMT in a normal rat tubular epithelial cell line (NRK52E). Activation of Smads by Ang II was attributed to degradation of an inhibitory Smad7, which was mediated by the AT1-Smurf2-dependent ubiquitin degradation mechanism because blockade of AT1 receptor or knockdown of Smurf2 inhibited Smad7 loss, thereby reducing Smad2/3 activation and EMT in response to ...

TY - JOUR. T1 - TGF-β coordinately activates TAK1/MEK/AKT/NFkB and SMAD pathways to promote osteoclast survival. AU - Gingery, Anne. AU - Bradley, Elizabeth W.. AU - Pederson, Larry. AU - Ruan, Ming. AU - Horwood, Nikki J.. AU - Oursler, Merry Jo. PY - 2008/9/10. Y1 - 2008/9/10. N2 - To better understand the roles of TGF-β in bone metabolism, we investigated osteoclast survival in response TGF-β and found that TGF-β inhibited apoptosis. We examined the receptors involved in promotion of osteoclast survival and found that the canonical TGF-β receptor complex is involved in the survival response. The upstream MEK kinase TAK1 was rapidly activated following TGF-β treatment. Since osteoclast survival involves MEK, AKT, and NFκB activation, we examined TGF-β effects on activation of these pathways and observed rapid phosphorylation of MEK, AKT, IKK, IκB, and NFκB. The timing of activation coincided with SMAD activation and dominant negative SMAD expression did not inhibit NFκB activation, ...

The occurrence and development of an endemic osteoarthritis Kashin-Beck Disease (KBD) is closely related to oxidative stress induced by free radicals. The aim of the study was to find the key signalling molecules or pathogenic factors as a potential treatment strategy for KBD.Real-time PCR and western blotting were performed to detect the mRNA and protein expression levels in cells and tissues. Immunohistochemical staining was assayed in rat models and human samples obtained from children. The type of cell death was identified by Annexin V and PI staining on a flow cytometry.Oxidative stress decreased levels of Smad2 and Smad3 in hypertrophic chondrocytes both in vitro and in vivo. In the cartilage of KBD patients, the expression of Smad2 and Smad3 proteins in the middle and deep zone were significantly decreased with an observed full deletion in the deep zone of some samples. Reduction of Smad2 protein induced necrotic death of hypertrophic chondrocytes, while reduction of Smad3 protein induced ...

The Spemann organizer induces neural tissue, dorsalizes mesoderm and generates a second dorsal axis. We report the isolation and characterization of Smad10, which has all three of these Spemann activities. Smad10 is expressed at the appropriate time to transduce Spemann signals endogenously. Like the organizer, Smad10 generates anterior and posterior neural tissues. Smad10 appears to function downstream of the Spemann organizer, consistent with a role in mediating organizer-derived signals. Interestingly, Smad10, unlike previously characterized mediators of Spemann activity, does not appear to block BMP signals. This finding, coupled with the functional activity and expression profile, suggests that Smad10 mediates Spemann action in a novel manner. ...

Results: Mice with fibroblast-specific Smad3 loss had accentuated adverse remodeling after reperfused infarction and exhibited an increased incidence of late rupture after nonreperfused infarction. The consequences of fibroblast-specific Smad3 loss were not a result of effects on acute infarct size but were associated with unrestrained fibroblast proliferation, impaired scar remodeling, reduced fibroblast-derived collagen synthesis, and perturbed alignment of myofibroblast arrays in the infarct. Polarized light microscopy in Sirius red-stained sections demonstrated that the changes in fibroblast morphology were associated with perturbed organization of the collagenous matrix in the infarcted area. In contrast, α-smooth muscle actin expression by infarct myofibroblasts was not affected by Smad3 loss. Smad3 critically regulated fibroblast function, activating integrin-mediated nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-2 (NOX-2) expression. Smad3 loss in cardiomyocytes attenuated ...

Since the initial description in 1986,29 numerous authors have described TGFβs ability to upregulate fibronectin synthesis in a host of different cell types. We began our studies by reconfirming that TGFβ upregulates the level of both fibronectin protein and mRNA in A10 aortic SMCs. Our findings were in accordance with those of others who have shown that the induction of fibronectin is a general response of vascular SMCs to TGFβ.. Although Smad proteins are well established as signal mediators downstream of TGFβ receptors, whether TGFβ stimulates fibronectin expression through a Smad-dependent pathway is controversial. Using various mutants of TGFβ type I receptor, Itoh et al showed that TGFβ-induced fibronectin expression requires activation of Smad proteins.30 Moreover, the authors demonstrated that the TGFβ-mediated fibronectin induction is absent in a Smad4-deficient cell line, MDA-MB-468, and this TGFβ dysfunction can be rescued by ectopic Smad4 expression. Here, in VSMCs, we ...

Bone morphogenetic proteins (BMPs) are members of the transforming growth factor (TGFβ) superfamily of secreted ligands that fulfil multiple functions during the development of vertebrate as well as non-vertebrate species (Hogan et al., 1996). Specific heteromeric type I/type II serine/threonine kinase receptor complexes mediate BMP action. Type I receptors, also termed ALKs (activin-receptor-like kinase) act downstream of type II receptors and determine the specificity within the receptor complex. The type I receptors, in turn, activate their intracellular downstream targets, known as Smads (Heldin et al., 1997). Smad proteins can be subdivided into three categories: receptor-regulated Smads (R-Smads), which transiently interact with type I receptors and become phosphorylated at their C-terminal motif SSXS; common partner Smads (Co-Smads), which associate with R-Smads and translocate them to the nucleus and modulate transcription of target genes; and inhibitory Smads (I-Smads), which can ...

SMAD4 is the only common SMAD in TGF-β signaling that usually impedes immune cell activation in the tumor microenvironment. However, we demonstrated here that selective deletion of Smad4 in NK cells actually led to dramatically reduced tumor cell rejection and augmented tumor cell metastases, reduced murine CMV clearance, as well as impeded NK cell homeostasis and maturation. This was associated with a downregulation of granzyme B (Gzmb), Kit, and Prdm1 in Smad4-deficient NK cells. We further unveiled the mechanism by which SMAD4 promotes Gzmb expression. Gzmb was identified as a direct target of a transcriptional complex formed by SMAD4 and JUNB. A JUNB binding site distinct from that for SMAD4 in the proximal Gzmb promoter was required for transcriptional activation by the SMAD4-JUNB complex. In a Tgfbr2 and Smad4 NK cell-specific double-conditional KO model, SMAD4-mediated events were found to be independent of canonical TGF-β signaling. Our study identifies and mechanistically ...

Chronic kidney disease (CKD) is a global socioeconomic problem. It is characterised by the presence of differentiated myofibroblasts that, in response to TGF B-1, produce tissue fibrosis, leading to renal failure. Here we define a novel interaction between the SET9 lysine methyltransferase and SMAD3, the principle mediator of TGF B-1 signalling in myofibroblasts. We show that SET9 deficient fibroblasts exhibit globally altered gene expression profiles in response to TGF B-1, whilst overexpression of SET9 enhances SMAD3 transcriptional activity. We also show that SET9 facilitates SMAD3 nuclear import and controls SMAD3 protein degradation, in a manner involving ubiquitination. On a cellular level, we demonstrate that SET9 is broadly required for TGF B-1 effects in diseased primary renal fibroblasts; SET9 promotes fibroblast migration into wounds, expression of extracellular matrix proteins, collagen contractility and myofibroblast differentiation. Finally, we demonstrate that SET9 is recruited to ...

Hepatocellular carcinoma (HCC) is certainly a main open public health problem credited to improved incidence, past due diagnosis and limited treatment options. in HCC sufferers. TGF- reliant cytostasis can be blunted in another group of cell lines (HLE, HLF, FLC-4) revealing high quantities of TGF- and Smad7 and displaying considerably decreased Smad3 Atrasentan hydrochloride signaling. Of those, HLF and HLE display past due TGF- signatures, which can be linked with poor treatment in HCC sufferers. RNAi with Smad3 blunted cytostatic results in PLC/PRF/5, HuH7 and Hep3B. HCC-M and HCC-T represent a third group of cell lines missing cytostatic TGF- signaling despite solid and extended Smad3 phosphorylation and low Smad7 and TGF- phrase. Inhibitory linker phosphorylation, as in HCC-T, may disrupt C-terminally phosphorylated Smad3 function. In overview, we assort 10 HCC cell lines in at least two groupings with respect to Atrasentan hydrochloride TGF- awareness. Cell lines reactive to the TGF- ...

TY - JOUR. T1 - Erbin inhibits transforming growth factor β signaling through a novel Smad-interacting domain. AU - Dai, Fangyan. AU - Chang, Chenbei. AU - Lin, Xia. AU - Dai, Penggao. AU - Mei, Lin. AU - Feng, Xin Hua. PY - 2007/9/1. Y1 - 2007/9/1. N2 - Smad proteins are critical intracellular signaling mediators for the transforming growth factor β (TGFβ) superfamily. Here, we report that Erbin (for ErbB2/Her2-interacting protein), which contains leucine-rich repeats and a PDZ (PSD-95/DLG/ZO-1) domain, interacts specifically with Smad3 and, to a lesser extent, with Smad2 through a novel Smad-interacting domain (SID) adjacent to its PDZ domain. Increased expression of Erbin does not affect the level of TGFβ-induced phosphorylation of Smad2/Smad3, but it physically sequesters Smad2/Smad3 from their association with Smad4 and hence negatively modulates TGFβ-dependent transcriptional responses and cell growth inhibition. An isoform of Erbin encoded by an alternatively spliced transcript in ...

In this video cells express the transcription factor SMAD2 in the green Channel and the FUCCI cell cycle indicators (see reference below) in the blue and red Channels. In this system, cells have blue nuclear signal (Cdt1-mCerulean3) in G1 and change to red (Geminin-mCherry) in late G1 to stay red for S, G2 and M phases. This allows us to quantify cell cycle progression at single cell level while measuring the activity of the transcription factor SMAD2. In this case, cells are growing under no stimulation, in 1% serum for 40 hours. Note cells transitioning from blue to red and how mitosis occurs in red cells to create 2 cells that will later be expressing the blue marker.. Reference: Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. ...

Signals mediated by the transforming growth factor-beta superfamily of growth factors have been implicated in thymic epithelial cell (TEC) differentiation, homeostasis, and function, but a direct reliance on these signals has not been established. Here we demonstrate that a block in canonical transforming growth factor-beta signaling by the loss of Smad4 expression in TECs leads to qualitative changes in TEC function and a progressively disorganized thymic microenvironment. Moreover, the number of thymus resident early T-lineage progenitors is severely reduced in the absence of Smad4 expression in TECs and directly correlates with extensive thymic and peripheral lymphopenia. Our observations hence place Smad4 within the signaling events in TECs that determine total thymus cellularity by controlling the number of early T-lineage progenitors.

With cardiovascular disease (CVD) being the leading cause of morbidity and death in the United States and worldwide (2, 28), studying the mechanisms of these pathologies is imperative. Recent studies have shown that a correlation exists between the activation of synthetic and growth-promoting transforming growth factor-β1 (TGF-β1) and the pathogenesis of CVD (13). Cell-to-cell adhesion through components of the extracellular matrix (ECM) is required for normal growth conditions; however, these adhesive interactions have also been linked to CVD pathogenesis. TGF-β1 is thought to synthesize ECM elements through a Smad3-dependent pathway. Considering the correlation between TGF-β1 and CVD, studying its mechanistic effects on cell proliferation and migration could prove beneficial in combatting CVD pathologies. Past studies involving TGF-β1 have generally focused on its intracellular Smad3 signals, and results have shown conflicting effects of Smad3 and even it switching between pro-growth and ...

Depending on the cellular context, transforming growth factor β (TGFβ) has been observed to exert protumorigenic functions, such as inducing an epithelial-mesenchymal transition (EMT), or inhibit tumorigenesis by activating apoptosis. The proapoptotic functions of TGFβ have been linked with the transcription factor SMAD4, which acts downstream of TGFβ and is frequently deleted in pancreatic ductal carcinoma (PDAC). To elucidate the mechanism by which TGFβ promotes apoptosis, David and colleagues used a Kras-mutant/Smad4-deleted PDAC murine model and found that reintroduction of SMAD4 sensitized cells to TGFβ treatment and promoted changes in cell morphology and loss of E-cadherin consistent with EMT. Moreover, SNAIL was shown to be upregulated following TGFβ treatment, and genetic depletion of SNAIL inhibited TGFβ-induced EMT, apoptosis, and accelerated pancreatic carcinogenesis in SMAD4-wild-type cells, raising the unexpected possibility that EMT precedes apoptosis and is required for ...

In this issue of JEM, Fu et al. described a novel mechanism in which reactive oxygen species (ROS)-activated Mink1 directly binds to and phosphorylates Smad2 at the inhibitory residue T324, preventing its nuclear localization (see figure). Consistent with its role in inhibiting Smad2 activation, cells lacking Mink1 displayed an opposite phenotype to Smad2-deficient cells (Malhotra et al., 2010; Martinez et al., 2010; Yoon et al., 2015). Not only TGF-β1 but also TGF-β3 stimulation in the presence of IL-6 results in enhanced Th17 cell generation in Mink1-deficient cells. Thus, it is plausible that Smad1, activated by TGF-β3 (Lee et al., 2012), could be directly phosphorylated and inhibited by Mink1 because it also contains this same inhibitory residue. Moreover, although both Smad1 and Smad2 possess the inhibitory T324 residue, Smad3 does not and is not phosphorylated by Mink1 (see figure). It is therefore possible that the inhibitory role of Mink1 in Th17 cell differentiation is not only ...

BMP‐4 signaling is known to inhibit neural induction in Xenopus (Jones et al., 1992; reviewed in Dale and Wardle, 1999). Interestingly, the BMP‐4 promoter contains ZEB sites and is itself BMP dependent (Vainio et al., 1993; Ebara et al., 1997; Kawasaki et al., 1999). As shown in Figure 7, we found that BMP‐4 expression was inhibited by ZEB‐2/SIP1, which could explain the neuralization/dorsalization effect of ZEB‐2/SIP1. This in vivo repression of Smad‐dependent genes by ZEB‐2/SIP1 is also consistent with the inhibition of Smad signaling by ZEB‐2/SIP1 that we observed in cell line systems, as described in this manuscript and in Postigo (2003).. Next, we examined the effect of ZEB‐1/δEF1 RNA injection into Xenopus embryos. Consistent with our results in cultured cell lines showing a synergy between TGFβ family members and ZEB‐1/δEF1, we found that microinjection of ZEB‐1/δEF1 also had an opposite effect to ZEB‐2/SIP1 in vivo, as it induced the ectopic expression of the ...

TGF-β is considered a master switch in the pathogenesis of organ fibrosis. The primary mediators of this activity are the SMAD proteins, particularly SMAD3. In the current study, we have developed a cell-penetrating peptide (CPP) conjugate of the HIV TAT protein that is fused to an aminoterminal sequence of sorting nexin 9 (SNX9), which was previously shown to bind phosphorylated SMAD3 (pSMAD3). We determined that specifically preventing the nuclear import of pSMAD3 using the TAT-SNX9 peptide inhibited profibrotic TGF-β activity in murine cells and human lung fibroblasts as well as in vivo with no demonstrable toxicity. TGF-β signaling mediated by pSMAD2, bone morphogenetic protein 4 (BMP4), EGF, or PDGF was unaffected by the TAT-SNX9 peptide. Furthermore, while the TAT-SNX9 peptide prevented TGF-βs profibrotic activity in vitro as well as in 2 murine treatment models of pulmonary fibrosis, a 3-amino acid point mutant that was unable to bind pSMAD3 proved ineffective. These findings ...

For the first time world-leading experts in the area of cellular signaling have joined to the production of a book on Smad signal transduction. Smads are the principal intracellular effectors of TGF-b

Fibrosis occurs when transforming growth factor-β (TGF-β) signaling is not attenuated after wound healing and fibroblasts remain activated, resulting in tissue scarring. The mechanisms that turn off TGF-β signaling after wound healing and inhibit this pathway in general are not well-understood. Palumbo-Zerr et al. noted that the abundance of the orphan receptor NR4A1 (nuclear receptor 4A1) was increased at the mRNA and protein levels in the fibrotic skin of patients with systemic sclerosis and in the skin of mice overexpressing a constitutively active TGF-β receptor type I (TBRI), which are a model for fibrosis. TGF-β stimulated the increased expression of NR4A1 through a complex containing the transcriptional effectors SMAD3, SMAD4, and SP1. Global Nr4a1 deficiency enhanced the expression of TGF-β target genes and fibrosis in mice overexpressing TBRI, as well as in other mouse models of fibrosis, including pulmonary fibrosis induced by bleomycin. Nr4a1-/- fibroblasts deposited more ...

Smad2 KO cell lysate available now. KO validated by Western Blot (WB). Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2 and 1 bp insertion in…

The KOMP Repository Collection is located at the MMRRC at the University of California, Davis and Childrens Hospital Oakland Research Institute. Question? Comments? For Mice, Cells, and germplasm please contact us at [email protected], US 1-888-KOMP-MICE or International +1-530-752-KOMP, or for vectors [email protected] or +1-510-450-7917 ...

The KOMP Repository is located at the University of California Davis and Childrens Hospital Oakland Research Institute. Question? Comments? For Mice, Cells, and germplasm please contact us at [email protected], US 1-888-KOMP-MICE or International +1-530-752-KOMP, or for vectors [email protected] or +1-510-450-7917 ...

SKI and SnoN proteins have been shown to inhibit TGF-β signaling, acting both as transcriptional co-repressors in the cell nucleus, and as sequestrators of SMAD

Detailed information about the celline expression of SMAD4 in Leukemia, B-ALL stained with HPA019154. The antibody showed a Not detected level of staining

The Hippo pathway plays a crucial role in growth control, proliferation and tumor suppression. Activity of the signaling pathway is associated with cell density sensing and tissue organization. Furthermore, the Hippo pathway helps to coordinate cellular processes through crosstalk with growth-factor-mediated signaling pathways such as TGF beta. Here we have examined the localization of interactions between proteins of the Hippo pathway (YAP/TAZ) and TGF beta (Smad2/3) signaling pathway by using in situ proximity ligation assays. We investigated the formation of protein complexes between YAP/TAZ and Smad2/3 and examined how these interactions were affected by TGF beta stimulation and cell density in HaCaT keratinocytes and in Smad4-deficient HT29 colon cancer cells. We demonstrate that TGF beta induces formation of YAP/TAZ-Smad2/3 complexes in HaCaT cells. Under sparse cell conditions, the complexes were detected to a higher degree and were predominantly located in the nucleus, while under dense ...

Kit contents: 1. MICROTITER PLATE * 1 2. ENZYME CONJUGATE*1 vial 3. STANDARD A*1 vial 4. STANDARD B*1 vial 5. STANDARD C*1 vial 6. STANDARD D*1 vial 7. STANDARD E*1 vial 8. STANDARD F*1 vial 9. SUBSTRATE A*1 vial 10. SUBSTRATE B*1 vial 11. STOP ...

Smad1 (phospho Ser465) antibody (SMAD family member 1) for WB. Anti-Smad1 (phospho Ser465) pAb (GTX50330) is tested in Human, Mouse, Rat samples. 100% Ab-Assurance.

SMAD5 antibody [N2C2], Internal (SMAD family member 5) for ICC/IF. Anti-SMAD5 pAb (GTX105101) is tested in Human samples. 100% Ab-Assurance.

Reactome is pathway database which provides intuitive bioinformatics tools for the visualisation, interpretation and analysis of pathway knowledge.

Although evidence indicating that COX-2 is involved in gastric carcinogenesis is accumulating (2, 3, 4) , the mechanisms responsible for the up-regulation of COX-2 are not completely understood. In the present study, we found that COX-2 is frequently overexpressed in tumors with increased expression levels of HER-2 or with reduced expression levels of Smad4 and that each of these proteins was associated with postoperative survival. Furthermore, a multivariate analysis demonstrated a significantly poor outcome in patients with Smad4-reduced tumors and concomitant COX-2 overexpression.. Several observations of the expression of COX-2 in human gastric cancer have been made using immunohistochemistry or Western blotting analyses (5, 6, 7, 8) . These data show a high frequency (58-68%) of COX-2 immunoreactivity, whereas the reported correlations between COX-2 expression and the clinicopathological characteristics and patient outcome were controversial. Lim et al. (32) did not find any significant ...

The lollipop plot above illustrates recurrent (observed in 3 or more out of 4440 TCGA tumor samples from 15 cancer types) and therefore potentially oncogenic missense mutations (click on Show Cancer Mutations). The bar plot below shows the proportion of tumor samples that have any kind of altering mutation(s) in the given protein. ...

Laboratory of Gene Regulation and Signal Transduction, Department of Pharmacology, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0723, USA. ...