Serine-Threonine Kinase Receptor-associated Protein Inhibits Apoptosis Signal-regulating Kinase 1 Function through Direct Interaction

Jung, Hye Young; Seong, Hyun‐A; Manoharan, Ravi; Ha, Hyunjung

doi:10.1074/jbc.m109.045229

Cited by 38 publications

(62 citation statements)

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“…Binding Assay, Immunoprecipitation, and Immunoblot-In vivo binding assays were performed as previously described (5,19). Native PAGE to determine the in vitro interaction between B-MYB and STRAP was performed using in vitro translated 35 S-labeled B-MYB, prepared with the TNT reticulocyte lysate system (Promega), and recombinant STRAP proteins (5).…”

Section: Methodsmentioning

confidence: 99%

“…STRAP also contributes to tumor progression by blocking TGF-␤-mediated signaling, especially in colon and lung carcinomas, indicating the possible involvement of STRAP in tumorigenesis (2,3). Recently, STRAP has been found to act as a regulator of 3-phosphoinositide-dependent protein kinase-1 (PDK1) and apoptosis signal-regulating kinase 1 (ASK1) to stimulate cell growth (4,5). In contrast to the findings described above, STRAP has also been shown to potentiate p53-induced apoptosis through direct interaction with p53 (6).…”

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confidence: 99%

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B-MYB Positively Regulates Serine-Threonine Kinase Receptor-associated Protein (STRAP) Activity through Direct Interaction

Seong

Manoharan

2011

Journal of Biological Chemistry

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Serine-threonine kinase receptor-associated protein (STRAP) functions as a regulator of both TGF-␤ and p53 signaling. However, the regulatory mechanism of STRAP activity is not understood. In this study, we report that B-MYB is a new STRAP-interacting protein, and that an amino-terminal DNAbinding domain and an area (amino acids 373-468) between the acidic and conserved regions of B-MYB mediate the B-MYB⅐STRAP interaction. Functionally, B-MYB enhances STRAP-mediated inhibition of TGF-␤ signaling pathways, such as apoptosis and growth inhibition, by modulating complex formation between the TGF-␤ receptor and SMAD3 or SMAD7. Furthermore, coexpression of B-MYB results in a dose-dependent increase in STRAP-mediated stimulation of p53-induced apoptosis and cell cycle arrest via direct interaction. Confocal microscopy showed that B-MYB prevents the normal translocation of SMAD3 in response to TGF-␤1 and stimulates p53 nuclear translocation. These results suggest that B-MYB acts as a positive regulator of STRAP.Serine-threonine kinase receptor-associated protein (STRAP) 2 is a transforming growth factor-␤ (TGF-␤) receptor-interacting protein that inhibits TGF-␤ signaling by stabilizing the association between TGF-␤ receptors and SMAD7 (1). STRAP also contributes to tumor progression by blocking TGF-␤-mediated signaling, especially in colon and lung carcinomas, indicating the possible involvement of STRAP in tumorigenesis (2, 3). Recently, STRAP has been found to act as a regulator of 3-phosphoinositide-dependent protein kinase-1 (PDK1) and apoptosis signal-regulating kinase 1 (ASK1) to stimulate cell growth (4, 5). In contrast to the findings described above, STRAP has also been shown to potentiate p53-induced apoptosis through direct interaction with p53 (6). These findings suggest that STRAP might have a bipartite role in the regulation of cell growth. However, the mechanism of STRAP regulation is not understood.B-myb is a member of the Myb family of transcription factors, which is ubiquitously expressed and involved in controlling cell proliferation and differentiation (7-9). B-myb is phylogenetically the most divergent of the three myb proteins, A-myb, B-myb, and c-myb (10). Numerous reports have established a critical role of B-myb in the growth of normal and tumorigenic cell lines (7,8,11,13). The most straightforward explanation for this effect is that B-myb could contribute to cell survival by inducing anti-apoptotic genes. For example, B-myb, like c-myb, stimulated transcription of Bcl2 and enhanced cell survival (15), suggesting that Bcl2 is a target gene for B-myb-mediated cell survival. In addition, anti-apoptotic ApoJ/Clusterin, which is highly induced in the presence of a variety of apoptotic stimuli, was also transactivated by B-MYB (16). Despite evidence supporting an anti-apoptotic function for B-myb, other studies have implicated B-myb in promoting cell death. Overexpression of B-myb accelerates apoptosis in TGF-␤1-treated M1 cells without affecting the regulation of c-myb and c-myc expression (17)....

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

B-MYB Positively Regulates Serine-Threonine Kinase Receptor-associated Protein (STRAP) Activity through Direct Interaction

Seong

Manoharan

2011

Journal of Biological Chemistry

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“…Whereas YWHAE interacts with Bad (89,90), PKC (91), and Ask1 (92), STRAP synergizes with Smad7 in the inhibition of TGF-␤ signaling, thus blocking apoptosis (93,94). Moreover, a novel inhibitory function for STRAP regarding Ask1-induced apoptosis has recently been proposed (95).…”

Section: Exploring Novel CC Protein Interaction Partners Using Amentioning

confidence: 99%

Structural and Functional Analysis of Novel Human Cytochrome c Targets in Apoptosis

Martínez-Fábregas

Dı́az-Moreno

González‐Arzola

et al. 2014

Molecular & Cellular Proteomics

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Since the first description of apoptosis four decades ago, great efforts have been made to elucidate, both in vivo and in vitro, the molecular mechanisms involved in its regulation. Although the role of cytochrome c during apoptosis is well established, relatively little is known about its participation in signaling pathways in vivo due to its essential role during respiration. To obtain a better understanding of the role of cytochrome c in the onset of apoptosis, we used a proteomic approach based on affinity chromatography with cytochrome c as bait in this study. In this approach, novel cytochrome c interaction partners were identified whose in vivo interaction and cellular localization were facilitated through bimolecular fluorescence complementation. Modeling of the complex interface between cytochrome c and its counterparts indicated the involvement of the surface surrounding the heme crevice of cytochrome c, in agreement with the vast majority of known redox adducts of cytochrome c. However, in contrast to the high turnover rate of the mitochondrial cytochrome c redox adducts, those occurring under apoptosis led to the formation of stable nucleo-cytoplasmic ensembles, as inferred mainly from surface plasmon resonance and nuclear magnetic resonance measurements, which permitted us to corroborate the formation of such complexes in vitro. The results obtained suggest that human cytochrome c interacts with pro-survival, anti-apoptotic proteins following its release into the cytoplasm. Thus, cytochrome c may interfere with cell survival pathways and unlock apoptosis in order to prevent the spatial and temporal coexistence of antagonist signals. Molecular & Cellular

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“…Alexa Fluor-594 anti-mouse and Alexa Fluor-488 anti-rabbit secondary antibodies were purchased from Molecular Probes Inc. (Eugene, OR). An inducible MPK38 shRNA NIH 3T3 cell line was generated using the following oligonucleotides: forward primer, 5Ј-TCGAGGCAGGCAGACAATGGAGGATTTCAAGAGA-ATCCTCCATTGTCTGCCTGCTTTTTTA-3Ј, containing an MPK38 sequence (underlined); and reverse primer, 5Ј-AGCT-TAAAAAAGCAGGCAGACAATGGAGGATTCTCTTGAA-ATCCTCCATTGTCTGCCTGCC-3Ј, containing an MPK38 sequence (underlined), as described previously (25). Inducible MPK38 shRNA stable clones were screened in the presence of 450 g/ml G418 for 14 days until all control parental NIH 3T3 cells completely died.…”

Section: Methodsmentioning

confidence: 99%

“…Construction of Smad Mutants-Smad mutants used for the in vitro kinase assays were generated by PCR under reaction parameters described previously (25). The following primers were used: for Smad2: forward (5Ј-GCGAATTCATGTC-GTCCATCTTGCCATTC-3Ј) and reverse (5Ј-GCCTCGAGT-TATGACATGCTTGAGCAACG-3Ј); for Smad3: forward (5Ј-GCGAATTCATGTCGTCATCCTGCCTTTC-3Ј) and reverse (5Ј-GCCTCGAGCTAAGACACACTGGAACAGCG-3Ј); for Smad4: forward (5Ј-GCGAATTCATGGACAATATGTCTA-TTACG-3Ј) and reverse (5Ј-GCGCCTCGAGTCAGTCTAA-AGGTTGTGGGTC-3Ј); for Smad7: forward (5Ј-GCGAA-TTCATGTTCAGGACCAAACGATCT-3Ј) and reverse (5Ј-GCCTCGAGCTACCGGCTGTTGAAGATGAC-3Ј).…”

Section: Methodsmentioning

confidence: 99%

Murine Protein Serine/Threonine Kinase 38 Stimulates TGF-β Signaling in a Kinase-dependent Manner via Direct Phosphorylation of Smad Proteins

Seong

Jung²,

2010

Journal of Biological Chemistry

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The present study demonstrated that murine protein serine/ threonine kinase 38 (MPK38) coimmunoprecipitates with Smad proteins (Smad2, -3, -4, and -7) and that this association is mediated by the catalytic kinase domain of MPK38. The association between MPK38 and Smad2, -3, and -4 was significantly increased by TGF-␤ or ASK1 signals, whereas these signals decreased association of MPK38 with Smad7. MPK38 stimulated TGF-␤-induced transcription required for TGF-␤-mediated biological functions, such as apoptosis and cell growth arrest, in a kinase-dependent manner. Knockdown of endogenous MPK38 showed an opposite effect, inhibiting TGF-␤ signaling. MPK38-mediated phosphorylation of Smad proteins (Ser 245 of Smad2, Ser 204 of Smad3, Ser 343 of Smad4, and Thr 96 of Smad7) was also found to be crucial to the positive regulation of TGF-␤ signaling induced by MPK38. In addition, MPK38 enhanced nuclear translocation of Smad3, as well as redistribution of Smad7 from the nucleus to the cytoplasm, in response to TGF-␤. Together, these results indicate that MPK38 functions as a stimulator of TGF-␤ signaling through direct interaction with and phosphorylation of Smad proteins.Type I transforming growth factor-␤ (TGF-␤) receptor kinases are serine/threonine kinases with roles in a wide array of cellular processes. Phosphorylation of the C-terminal SXS motif in receptor-regulated Smads (R-Smads) by these kinases is a crucial step in TGF-␤ family signaling (1-3). R-Smad phosphorylation results in the formation of heterodimeric complexes with the common Smad, Smad4. In addition, the phosphorylation of serine or threonine residues in the N-terminal or linker region has been observed in endogenous R-Smads (4). Several lines of evidence have demonstrated the existence of kinases responsible for N-terminal or linker region phosphorylation events, including mitogen-activated protein kinases (MAPKs), Ca 2ϩ and calmodulin-dependent kinase II, cyclindependent kinase (CDK), 3 protein kinase C (PKC), and G protein-coupled receptor kinase 2 (5). Common Smad, Smad4, is constitutively phosphorylated in cells, although not all of the phosphorylation sites are known. Extracellular signal-regulated kinase (ERK) has been shown to phosphorylate Smad4 at Thr 277 , resulting in the prevention of nuclear translocation of Smad4 (6). The inhibitory Smads (I-Smads), Smad6 and Smad7, are phosphorylated by uncharacterized kinases (7,8). This provides a mechanism for integration of the Smad pathway with other signaling pathways, in which kinases act as essential mediators, which modulate Smad-mediated signaling. Thus, the identification and characterization of additional cellular kinases responsible for Smad phosphorylation could lead to a better understanding of the regulatory role of Smad phosphorylations, especially in the N-terminal and linker region, in TGF-␤ signaling.Murine protein serine/threonine kinase 38 (MPK38), also known as pEg3 kinase and maternal embryonic leucine zipper kinase (Melk), is a member of the AMP-activated protein kinase family o...

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Serine-Threonine Kinase Receptor-associated Protein Inhibits Apoptosis Signal-regulating Kinase 1 Function through Direct Interaction

Cited by 38 publications

References 33 publications

B-MYB Positively Regulates Serine-Threonine Kinase Receptor-associated Protein (STRAP) Activity through Direct Interaction

B-MYB Positively Regulates Serine-Threonine Kinase Receptor-associated Protein (STRAP) Activity through Direct Interaction

Structural and Functional Analysis of Novel Human Cytochrome c Targets in Apoptosis

Murine Protein Serine/Threonine Kinase 38 Stimulates TGF-β Signaling in a Kinase-dependent Manner via Direct Phosphorylation of Smad Proteins

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