Hematopoietic stem cells (HSC) self-renew to sustain stem cell pools and differentiate to generate all types of blood cells. HSCs remain in quiescence to sustain their long-term self-renewal potential. It remains unclear whether protein... more
Hematopoietic stem cells (HSC) self-renew to sustain stem cell pools and differentiate to generate all types of blood cells. HSCs remain in quiescence to sustain their long-term self-renewal potential. It remains unclear whether protein quality control is required for stem cells in quiescence when RNA content, protein synthesis, and metabolic activities are profoundly reduced. Here, we report that protein quality control via endoplasmic reticulum-associated degradation (ERAD) governs the function of quiescent HSCs. The Sel1L/Hrd1 ERAD genes are enriched in the quiescent and inactive HSCs, and conditional knockout of Sel1L in hematopoietic tissues drives HSCs to hyperproliferation, which leads to complete loss of HSC self-renewal and HSC depletion. Mechanistically, ERAD deficiency via Sel1L knockout leads to activation of mammalian target of rapamycin (mTOR) signaling. Furthermore, we identify Ras homolog enriched in brain (Rheb), an activator of mTOR, as a novel protein substrate of...
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AMPK ensures cell survival during energetic stress by stimulating the kinase Akt through mTORC2.
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Research Interests: Genetics, Nephrology, Biology, Dialysis, Enzyme Inhibitors, and 15 moreMedicine, Gene expression, Kidney, Animals, Phosphorylation, ERK, Mitogen Activated Protein Kinase, Clinical Sciences, mRna expression levels, DNA binding, Antineoplastic Agents, Mesangial Cells, Plasminogen Activator Inhibitor, Interferon gamma, and Kidney glomerulus
Research Interests: Mass Spectrometry, Biology, Biological Chemistry, Medicine, Biological Sciences, and 15 moreHumans, Protein Kinases, Phosphorylation, Proteins, Plasmids, CHEMICAL SCIENCES, Time Factors, Pyridines, Glutathione Transferase, Imidazoles, Amino Acid Sequence, Protein Binding, Molecular Sequence Data, binding sites, and Medical and Health Sciences
Approximately 35% of follicular thyroid carcinomas and a small fraction of follicular adenomas are associated with a t(2;3)(q13;p25) chromosomal translocation that fuses paired box gene 8 (PAX8) with the peroxisome proliferator-activated... more
Approximately 35% of follicular thyroid carcinomas and a small fraction of follicular adenomas are associated with a t(2;3)(q13;p25) chromosomal translocation that fuses paired box gene 8 (PAX8) with the peroxisome proliferator-activated receptor-γ gene (PPARG), resulting in expression of a PAX8-PPARγ fusion protein, PPFP. The mechanism by which PPFP contributes to follicular thyroid neoplasia is poorly understood. Therefore, we have created mice with thyroid-specific expression of PPFP. At 1 yr of age, 25% of PPFP mice demonstrate mild thyroid hyperplasia. We bred these mice to mice with thyroid-specific single-allele deletion of the tumor suppressor Pten, denoted ThyPten+/−. In humans, PTEN deletion is associated with follicular adenomas and carcinomas, and in mice, deletion of one Pten allele causes mild thyroid hyperplasia. We found that PPFP synergizes with ThyPten+/− to cause marked thyroid hyperplasia, but carcinomas were not observed. AKT phosphorylation was increased as exp...
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Research Interests: Mixed Methods, Flow Cytometry, Fluorescence Microscopy, Biology, Health, and 15 moreAutophagy, Apoptosis, Medicine, Cell Culture, Biological Sciences, Humans, Animals, Cell Death, ER stress, Knowledge base, Eukaryotic Cells, Human Disease, Biological Assay, Biochemistry and cell biology, and Immunoblotting
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These include: This article cites 59 articles, 22 of which can be accessed free at:
ABSTRACTAlthough visceral adipocytes located within the body’s central core are maintained at ~37°C, adipocytes within bone marrow, subcutaneous, and dermal depots are found primarily within the peripheral shell, and generally exist at... more
ABSTRACTAlthough visceral adipocytes located within the body’s central core are maintained at ~37°C, adipocytes within bone marrow, subcutaneous, and dermal depots are found primarily within the peripheral shell, and generally exist at cooler temperatures. Responses of brown and beige/brite adipocytes to cold stress are well-studied; however, comparatively little is known about mechanisms by white adipocytes adapt to temperatures below 37°C. Here we report that adaptation of cultured adipocytes to 31°C, the temperature at which distal marrow adipose tissues and subcutaneous adipose tissues often reside, induces extensive changes in gene expression, increased anabolic and catabolic lipid metabolism, and elevated oxygen consumption with reduced reliance on glucose and preferential use of pyruvate, glutamine and fatty acids as energy sources. Cool temperatures up-regulate stearoyl-CoA desaturase-1 expression and monounsaturated lipid levels in cultured adipocytes and distal bone marrow...
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The mechanistic target of rapamycin complex 1 (mTORC1) is a major serine/threonine kinase that stimulates cellular anabolic processes including protein and lipid synthesis while suppressing catabolic processes such as autophagy in... more
The mechanistic target of rapamycin complex 1 (mTORC1) is a major serine/threonine kinase that stimulates cellular anabolic processes including protein and lipid synthesis while suppressing catabolic processes such as autophagy in response to growth factors and amino acids (1). Upon mTORC1 activation, it phosphorylates multiple substrates including S6 kinase (S6K), eIF4E binding protein (4EBP), and unc-51-like kinase (ULK) (2). Both S6K and 4EBP are key regulators for mRNA translation and cell cycle progression (3). In addition to mTORC1’s roles in stimulating these anabolic processes, mTORC1-dependent ULK phosphorylation inhibits its kinase activity, which is essential for autophagy induction (4). Thus, mTORC1 activation in response to growth factors and amino acids promotes key cellular anabolic processes while it suppresses major catabolic processes, to build biosynthetic molecules essential for cell growth and proliferation.
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Tsukasa Suzuki,1 Dave Bridges,1 Daisuke Nakada,1,7 Georgios Skiniotis,1,3 Sean J. Morrison,1,2,8 Jiandie D. Lin,1,4 Alan R. Saltiel,1,5,6 and Ken Inoki1,5,6,* 1Life Sciences Institute 2Howard Hughes Medical Institute University of... more
Tsukasa Suzuki,1 Dave Bridges,1 Daisuke Nakada,1,7 Georgios Skiniotis,1,3 Sean J. Morrison,1,2,8 Jiandie D. Lin,1,4 Alan R. Saltiel,1,5,6 and Ken Inoki1,5,6,* 1Life Sciences Institute 2Howard Hughes Medical Institute University of Michigan, Ann Arbor, MI 48109, USA 3Department of Biological Chemistry 4Department of Cell and Developmental Biology 5Department of Molecular and Integrative Physiology 6Department of Internal Medicine University of Michigan Medical School, Ann Arbor, MI 48109, USA 7Present address: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USA 8Present address: Children’s Research Institute, Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA *Correspondence: inokik@umich.edu http://dx.doi.org/10.1016/j.molcel.2013.03.022
Glycolysis is upregulated under conditions such as hypoxia and high energy demand to promote cell proliferation, although the mechanism remains poorly understood. We find that hypoxia in Saccharomyces cerevisiae induces concentration of... more
Glycolysis is upregulated under conditions such as hypoxia and high energy demand to promote cell proliferation, although the mechanism remains poorly understood. We find that hypoxia in Saccharomyces cerevisiae induces concentration of glycolytic enzymes, including the Pfk2p subunit of the rate-limiting phosphofructokinase, into a single, non-membrane-bound granule termed the "glycolytic body" or "G body." A yeast kinome screen identifies the yeast ortholog of AMP-activated protein kinase, Snf1p, as necessary for G-body formation. Many G-body components identified by proteomics are required for G-body integrity. Cells incapable of forming G bodies in hypoxia display abnormal cell division and produce inviable daughter cells. Conversely, cells with G bodies show increased glucose consumption and decreased levels of glycolytic intermediates. Importantly, G bodies form in human hepatocarcinoma cells in hypoxia. Together, our results suggest that G body formation is...
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The RNA binding protein, LARP1, has been proposed to function downstream of mTORC1 to regulate the translation of 5'TOP mRNAs such as those encoding ribosome proteins (RP). However, the roles of LARP1 in the translation of 5'TOP... more
The RNA binding protein, LARP1, has been proposed to function downstream of mTORC1 to regulate the translation of 5'TOP mRNAs such as those encoding ribosome proteins (RP). However, the roles of LARP1 in the translation of 5'TOP mRNAs are controversial and its regulatory roles in mTORC1-mediated translation remain unclear. Here we show that LARP1 is a direct substrate of mTORC1 and Akt/S6K1. Deep sequencing of LARP1-bound mRNAs reveal that non-phosphorylated LARP1 interacts with both 5' and 3'UTRs of RP mRNAs and inhibits their translation. Importantly, phosphorylation of LARP1 by mTORC1 and Akt/S6K1 dissociates it from 5'UTRs and relieves its inhibitory activity on RP mRNA translation. Concomitantly, phosphorylated LARP1 scaffolds mTORC1 on the 3'UTRs of translationally-competent RP mRNAs to facilitate mTORC1-dependent induction of translation initiation. Thus, in response to cellular mTOR activity, LARP1 serves as a phosphorylation-sensitive molecular switc...
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The present invention relates to abnormalities in TSC signaling for identifying the compositions and methods of the conductive pathway. In particular, the present invention relates to a method such as tuberous sclerosis disorder diagnosis... more
The present invention relates to abnormalities in TSC signaling for identifying the compositions and methods of the conductive pathway. In particular, the present invention relates to a method such as tuberous sclerosis disorder diagnosis and treatment, the disorder is caused by mutations in the TSC genes of. The present invention further relates to methods and compositions for treating cancer signal transduction by the TSC disorder mediated.
Tuberous sclerosis complex (TSC) is a genetic disease caused by a mutation in either the tsc1 or tsc2 tumor suppressor gene. Recent studies have demonstrated that TSC2 displays GAP (GTPase-activating protein) activity specifically towards... more
Tuberous sclerosis complex (TSC) is a genetic disease caused by a mutation in either the tsc1 or tsc2 tumor suppressor gene. Recent studies have demonstrated that TSC2 displays GAP (GTPase-activating protein) activity specifically towards the small G protein Rheb and inhibits its ability to stimulate the mTOR signaling pathway. Rheb and TSC2 comprise a unique pair of GTPase and GAP, because Rheb has high basal GTP levels and TSC2 does not have the catalytic arginine finger found in Ras-GAP. To investigate the function of TSC2 and Rheb in mTOR signaling, we analyzed the TSC2-stimulated Rheb GTPase activity. We found that Arg15, a residue equivalent to Gly12 in Ras, is important for Rheb to function as a substrate for TSC2 GAP. In addition, we identified asparagine residues essential for TSC2 GAP activity. We demonstrated a novel catalytic mechanism of the TSC2 GAP and Rheb that TSC2 uses a catalytic “asparagine thumb” instead of the arginine finger found in Ras-GAP. Furthermore, we d...
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This study sought to determine whether platelet-derived growth factor (PDGF) and angiotensin II (AngII) stimulate migration of cultured rat glomerular mesangial cells. After finding that this was so, the effects of adrenomedullin (ADM)... more
This study sought to determine whether platelet-derived growth factor (PDGF) and angiotensin II (AngII) stimulate migration of cultured rat glomerular mesangial cells. After finding that this was so, the effects of adrenomedullin (ADM) and cAMP-elevating agents on basal and stimulated mesangial cell migration were examined. Two isoforms of PDGF, AB and BB, stimulated migration in a concentration-dependent manner between 1 and 50 ng/ml, while the AA isoform lacked significant effect. AngII modestly but significantly stimulated migration in a concentration-dependent manner between 10(-7) and 10(-6) mol/L. Rat ADM significantly inhibited the PDGF BB- and AngII-stimulated migration in a concentration-dependent manner between 10(-8) and 10(-7) mol/L. Inhibition by rat ADM was accompanied by an increase in cellular cAMP. cAMP agonists or inducers such as 8-bromo cAMP, forskolin, and prostaglandin I2 also significantly reduced the stimulated migration. H 89, a protein kinase A (PKA) inhibi...
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Background / Purpose: Protein synthesis and autophagic degradation are regulated in an opposite manner by mammalian target of rapamycin (mTOR), whereas under certain conditions it would be beneficial if they occurred in unison to handle... more
Background / Purpose: Protein synthesis and autophagic degradation are regulated in an opposite manner by mammalian target of rapamycin (mTOR), whereas under certain conditions it would be beneficial if they occurred in unison to handle rapid protein turnover. Main conclusion: We observed a distinct cellular compartment at the trans side of the Golgi apparatus, the TOR-autophagy spatial coupling compartment (TASCC), where (auto)lysosomes and mTOR accumulated during Ras-induced senescence. mTOR recruitment to the TASCC was amino acid- and Rag guanosine triphosphatase-dependent, and disruption of mTOR localization to the TASCC suppressed interleukin-6/8 synthesis.TASCC formation was observed during macrophage differentiation and in glomerular podocytes; both displayed increased protein secretion. The spatial coupling of cells' catabolic and anabolic machinery could augment their respective functions and facilitate the mass synthesis of secretory proteins.
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Tuberous sclerosis complex (TSC) is a genetic disease caused by a mutation in either the tsc1 or tsc2 tumor suppressor gene. Recent studies have demonstrated that TSC2 displays GAP (GTPase-activating protein) activity specifically towards... more
Tuberous sclerosis complex (TSC) is a genetic disease caused by a mutation in either the tsc1 or tsc2 tumor suppressor gene. Recent studies have demonstrated that TSC2 displays GAP (GTPase-activating protein) activity specifically towards the small G protein Rheb and inhibits its ability to stimulate the mTOR signaling pathway. Rheb and TSC2 comprise a unique pair of GTPase and GAP, because Rheb has high basal GTP levels and TSC2 does not have the catalytic arginine finger found in Ras-GAP. To investigate the function of TSC2 and Rheb in mTOR signaling, we analyzed the TSC2-stimulated Rheb GTPase activity. We found that Arg15, a residue equivalent to Gly12 in Ras, is important for Rheb to function as a substrate for TSC2 GAP. In addition, we identified asparagine residues essential for TSC2 GAP activity. We demonstrated a novel catalytic mechanism of the TSC2 GAP and Rheb that TSC2 uses a catalytic "asparagine thumb" instead of the arginine finger found in Ras-GAP. Further...
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Although AMP-activated protein kinase (AMPK) is involved in regulating carbohydrate and lipid metabolism, activated AMPK also plays an anti-inflammatory role in many cell populations. However, despite the ability of AMPK activation to... more
Although AMP-activated protein kinase (AMPK) is involved in regulating carbohydrate and lipid metabolism, activated AMPK also plays an anti-inflammatory role in many cell populations. However, despite the ability of AMPK activation to diminish the severity of inflammatory responses, previous studies have found that AMPK activity is diminished in LPS-treated neutrophils and also in lungs of mice with LPS-induced acute lung injury (ALI). Since GSK3β participates in regulating AMPK activity, we examined potential roles for GSK3β in modulating LPS-induced activation of neutrophils and macrophages and in influencing severity of ALI. We found that GSK3β-dependent phosphorylation of T479-AMPK was associated with pT172 dephosphorylation and inactivation of AMPK following TLR4 engagement. GSK3β inhibitors BIO (6-bromoindirubin-3'-oxime), SB216763, or siRNA knockdown of GSK3β, but not the PI3K/AKT inhibitor LY294002, prevented Thr172-AMPK dephosphorylation. Exposure to LPS resulted in rap...
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mTOR, an evolutionarily conserved Ser/Thr protein kinase, belongs to the PI3K-related kinase family, which also includes DNA-PKcs, ATM, and ATR. Although other PI3K-related kinase family members have been shown to secure genomic integrity... more
mTOR, an evolutionarily conserved Ser/Thr protein kinase, belongs to the PI3K-related kinase family, which also includes DNA-PKcs, ATM, and ATR. Although other PI3K-related kinase family members have been shown to secure genomic integrity by sensing DNA damage and related stresses, mTOR is known to function as a nutrient and growth factor sensor. mTOR is the catalytic subunit of two distinct multiprotein complexes known as mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). In response to growth factor and nutrient availability, these complexes regulate a variety of cellular processes, such as cell growth, proliferation, and survival by modulating downstream effectors, such as S6K1, 4EBP1, and AKT. Therefore, evaluation of mTOR activity has been a clear readout in order to monitor the physiological status of cells in response to environmental cues. Here, we present the current techniques for the assessment of mTOR kinase activity in different experimental settings.
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Mammalian target of rapamycin (mTOR) is an evolutionarily conserved serine/threonine protein kinase implicated in a wide array of cellular processes such as cell growth, proliferation, and survival. Analogous to the situation in yeast,... more
Mammalian target of rapamycin (mTOR) is an evolutionarily conserved serine/threonine protein kinase implicated in a wide array of cellular processes such as cell growth, proliferation, and survival. Analogous to the situation in yeast, mTOR forms two distinct functional complexes termed mTOR complex 1 and 2 (mTORC1 and mTORC2). mTORC1 activity is inhibited by rapamycin, a specific inhibitor of mTOR, whereas mTORC2 activity is resistant to short-term treatments with rapamycin. In response to growth factors, mTORC2 phosphorylates Akt, an essential kinase involved in cell survival. On the other hand, mTORC1 can be activated by both growth factors and nutrients such as glucose and amino acids. In turn, mTORC1 regulates the activity of the translational machinery by modulating S6 kinase (S6K) activity and eIF4E binding protein 1 (4E-BP1) through direct phosphorylation. Consequently, protein synthesis and cell growth are stimulated in a variety of different cell types. In addition, mTORC1 inhibits autophagy, an essential protein degradation and recycling system, which cells employ to sustain their viability in times of limited availability of nutrients. Recent studies have highlighted the fact that autophagy plays crucial roles in many aspects of human health including cancer development, neurodegenerative disease, diabetes, and aging. It is likely that dysregulation of the mTOR-autophagy pathway may contribute at least in part to these human disorders. Therefore, the assessment of mTOR activity is important to understand the status of autophagy in the cells being analyzed and its role in autophagy-related disorders. In this section, we describe methods to monitor mTOR activity both in vitro and in vivo.
Research Interests: Biology, Immunohistochemistry, Autophagy, Medicine, Protein synthesis, and 14 moreCell line, Humans, Protein Kinases, Human health, Protein Degradation, Neurodegenerative Disease, Protein Kinase, Amino Acid Sequence, Growth Factor, Protein Binding, Cell Survival, Biochemistry and cell biology, Cell Growth, and In Vitro Techniques
LST8 is a component of both mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). Herein, to examine the role of LST8, a common component of mTOR complexes, in the regulation of mTORC1 and mTORC2, first, we showed over-expression of LST8... more
LST8 is a component of both mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). Herein, to examine the role of LST8, a common component of mTOR complexes, in the regulation of mTORC1 and mTORC2, first, we showed over-expression of LST8 in HepG2 to markedly enhance basal phosphorylation levels of not only p70 S6 kinase but also Akt. In contrast, LST8 knockdown by siRNA in HepG2 decreased phosphorylation levels of both p70 S6 kinase and Akt. These results indicate the LST8 expression level to determine basal mTORC1 and mTORC2 activities, since LST8 appears to be the component present at the lowest level in both mTORC1 and mTORC2 complexes. Previously, we reported S6 kinase phosphorylation to be reduced by over-expression of the Cterminally deleted Raptor mutant (Raptor-ΔCT) not binding to mTOR or LST8, while phosphorylation levels of Akt were markedly enhanced with no alteration in IRS-1 phosphorylation or PI 3-kinase activity. Using Raptor-ΔCT, we investigated the competition for association with LST8 between mTORC1 and mTORC2. Over-expression of Raptor-ΔCT abolished formation of the Raptor, S6 kinase, mTOR and LST8 complex, while the amount of LST8 in the Rictor-mTOR complex was increased. Therefore, it is likely that Raptor-mTOR and Rictor-mTOR complexes compete for association with LST8, and this mechanism may contribute to the reciprocal negative regulations of mTORC1 and mTORC2 activities, in terms of their LST8 components.:
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Tuberous sclerosis complex (TSC) is a genetic disease caused by mutation in either the tsc1 or tsc2 tumor suppressor genes. TSC1 and TSC2 protein form a physical and functional complex in vivo. Recent studies have demonstrated that TSC2... more
Tuberous sclerosis complex (TSC) is a genetic disease caused by mutation in either the tsc1 or tsc2 tumor suppressor genes. TSC1 and TSC2 protein form a physical and functional complex in vivo. Recent studies have demonstrated that TSC2 displays GTPase activating protein (GAP) activity specifically toward the small G protein Rheb (Ras homolog enriched in brain) and inhibits its ability to stimulate the mammalian target of rapamycin (mTOR) signaling pathway. We have presented three methods to determine the activity of TSC2 as a GAP toward the Rheb GTPase. The first involves the isolation of TSC2 from cells and measurement of its activity toward Rheb substrate in vitro. The second involves the measurement of Rheb-associated guanine nucleotides as measure of TSC2 GAP activity on Rheb in vivo. The last method is to determine the phosphorylation of S6K1 (ribosomal S6 kinase), which is a downstream target of mTOR, as an indirect assay for TSC2 GAP activity in vivo.
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The clock protein BMAL1 (brain and muscle Arnt-like protein 1) participates in circadian regulation of lipid metabolism, but its contribution to insulin AKT-regulated hepatic lipid synthesis is unclear. Here we used both Bmal1(-/-) and... more
The clock protein BMAL1 (brain and muscle Arnt-like protein 1) participates in circadian regulation of lipid metabolism, but its contribution to insulin AKT-regulated hepatic lipid synthesis is unclear. Here we used both Bmal1(-/-) and acute liver-specific Bmal1-depleted mice to study the role of BMAL1 in refeeding-induced de novo lipogenesis in the liver. Both global deficiency and acute hepatic depletion of Bmal1 reduced lipogenic gene expression in the liver upon refeeding. Conversely, Bmal1 overexpression in mouse liver by adenovirus was sufficient to elevate the levels of mRNA of lipogenic enzymes. Bmal1(-/-) primary mouse hepatocytes displayed decreased levels of de novo lipogenesis and lipogenic enzymes, supporting the notion that BMAL1 regulates lipid synthesis in hepatocytes in a cell-autonomous manner. Both refed mouse liver and insulin-treated primary mouse hepatocytes showed impaired AKT activation in the case of either Bmal1 deficiency or Bmal1 depletion by adenoviral s...
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The tumor-suppressor proteins TSC1 and TSC2 are associated with an autosomal dominant disorder known as tuberous sclerosis complex (TSC). TSC1 and TSC2 function as a heterodimer to inhibit cell growth and proliferation. Another protein,... more
The tumor-suppressor proteins TSC1 and TSC2 are associated with an autosomal dominant disorder known as tuberous sclerosis complex (TSC). TSC1 and TSC2 function as a heterodimer to inhibit cell growth and proliferation. Another protein, mTOR (mammalian target of rapamycin), is regarded as a central controller of cell growth in response to growth factors, cellular energy and nutrient levels. Recent breakthroughs in TSC research link the TSC1/2 heterodimer protein to the mTOR signaling network. It has recently been shown that TSC2 has GTPase-activating protein (GAP) activity towards the Ras family small GTPase Rheb (Ras homolog enriched in brain), and TSC1/2 antagonizes the mTOR signaling pathway via stimulation of GTP hydrolysis of Rheb. Thus, TSC1/2 and Rheb have pivotal roles in mediating growth factors, nutrient and energy sensing signals to mTOR-dependent targets. These discoveries lend new insight into TSC pathogenesis.
Research Interests: Biology, Medicine, Signal Transduction, Biological Sciences, Neuropeptides, and 12 moreHumans, Protein Kinases, Animals, CHEMICAL SCIENCES, Growth Factor, Protein Kinase Inhibitors, Cell Growth, Tuberous sclerosis complex, Autosomal Dominant, Signaling pathway, tumor suppressor, and Medical and Health Sciences
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Cells possess adaptive biosynthetic systems to maintain cellular energy levels for survival under adverse environmental conditions. Autophagy is an evolutionarily conserved cellular catabolic process that breaks down and recycles... more
Cells possess adaptive biosynthetic systems to maintain cellular energy levels for survival under adverse environmental conditions. Autophagy is an evolutionarily conserved cellular catabolic process that breaks down and recycles cytosolic material including macromolecules and organelles through lysosomal degradation. This catabolic process, represented by macroautophagy, is induced by a variety of cellular stresses such as nutrient starvation, which causes a shortage of cellular energy for cells to maintain cellular homeostasis and essential biological activities. In contrast, upon nutrient availability, cells stimulate anabolic processes. The mechanistic/mammalian target of rapamycin, a serine/threonine protein kinase, is a key player in stimulating cellular anabolism in response to nutrients and growth factors, and plays a crucial role in suppressing autophagy activity. Growing evidence has suggested that autophagy activity is required for the maintenance and physiological functions of renal cells including proximal tubular cells and podocytes. In this article, we discuss recent progress in the regulation of autophagy by mechanistic/mammalian target of rapamycin signaling.
Research Interests: Biology, Autophagy, Medicine, Signal Transduction, Humans, and 4 moreKidney, mTOR, Animals, and Clinical Sciences
Combining classic biochemical approaches with dynamic mathematical modeling elucidates a new model for insulin-mediated activation of mTORC2.
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A kinase complex associated with cellular growth has an unexpected role in muscle atrophy.
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A cellular compartment allows simultaneous protein synthesis and degradation.
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Research Interests: Biology, Autophagy, Medicine, Protein synthesis, Biological Sciences, and 13 moreHumans, Protein Kinases, Tumor Suppressor Gene, Tuberous sclerosis, Hamartoma, Human Disease, Growth Factor, Protein Binding, Cell Growth, Tuberous sclerosis complex, Cardiomegaly, Signaling pathway, and Medical and Health Sciences
Tuberous sclerosis (TSC) is an autosomal dominant disorder characterized by the formation of hamartomas in a wide range of human tissues. Mutation in either the TSC1 or TSC2 tumour suppressor gene is responsible for both the familial and... more
Tuberous sclerosis (TSC) is an autosomal dominant disorder characterized by the formation of hamartomas in a wide range of human tissues. Mutation in either the TSC1 or TSC2 tumour suppressor gene is responsible for both the familial and sporadic forms of this disease. TSC1 and TSC2 proteins form a physical and functional complex in vivo. Here, we show that TSC1-TSC2 inhibits the p70 ribosomal protein S6 kinase 1 (an activator of translation) and activates the eukaryotic initiation factor 4E binding protein 1 (4E-BP1, an inhibitor of translational initiation). These functions of TSC1-TSC2 are mediated by inhibition of the mammalian target of rapamycin (mTOR). Furthermore, TSC2 is directly phosphorylated by Akt, which is involved in stimulating cell growth and is activated by growth stimulating signals, such as insulin. TSC2 is inactivated by Akt-dependent phosphorylation, which destabilizes TSC2 and disrupts its interaction with TSC1. Our data indicate a molecular mechanism for TSC2 in insulin signalling, tumour suppressor functions and in the inhibition of cell growth.
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The kinase complex mechanistic target of rapamycin 1 (mTORC1) plays an important role in controlling growth and metabolism. We report here that the stepwise formation of phosphatidylinositol 3-phosphate (PI(3)P) and phosphatidylinositol... more
The kinase complex mechanistic target of rapamycin 1 (mTORC1) plays an important role in controlling growth and metabolism. We report here that the stepwise formation of phosphatidylinositol 3-phosphate (PI(3)P) and phosphatidylinositol 3,5-bisphosphate (PI(3,5)P2) regulates the cell type–specific activation and localization of mTORC1. PI(3)P formation depends on the class II phosphatidylinositol 3-kinase (PI3K) PI3K-C2α, as well as the class III PI3K Vps34, while PI(3,5)P2 requires the phosphatidylinositol-3-phosphate-5-kinase PIKFYVE. In this paper, we show that PIKFYVE and PI3K-C2α are necessary for activation of mTORC1 and its translocation to the plasma membrane in 3T3-L1 adipocytes. Furthermore, the mTORC1 component Raptor directly interacts with PI(3,5)P2. Together these results suggest that PI(3,5)P2 is an essential mTORC1 regulator that defines the localization of the complex.
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Oral mucosa progenitor/stem cells reside as a small-sized cell population that eventually differentiates concurrently with an increase in cell size. Activation of the mammalian target of rapamycin (mTOR) leads to an increase in cell size.... more
Oral mucosa progenitor/stem cells reside as a small-sized cell population that eventually differentiates concurrently with an increase in cell size. Activation of the mammalian target of rapamycin (mTOR) leads to an increase in cell size. We hypothesized that rapamycin, a specific inhibitor of mTOR, will maintain primary human oral keratinocytes as a small-sized, undifferentiated cell population capable of retaining their proliferative capacity. Primary, rapamycin-treated (2 nM, 20 nM) oral keratinocytes showed a diminished cell size that correlated with a higher clonogenicity, a longer-term proliferative potential, and a slower cycling cell population concurrent with decreased expression of a differentiation marker when compared with untreated cells. Only the 2-nM rapamycin-treated oral keratinocytes maintained their ability to regenerate oral mucosa in vitro after 15 weeks of culture. Rapamycin, a Food and Drug Administration-approved drug, may have applicability for use in creati...
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Research Interests: Biology, Calcium, Flavonoids, Gene expression, Endothelial Cells, and 15 moreCardiovascular Pharmacology, Aldosterone, Animals, Male, Animal Model, Cyclosporine, Clinical Sciences, Endothelial cell, Isoenzymes, Base Sequence, DNA binding proteins, Enzyme Induction, Immunosuppressive Agents, Drug synergism, and Cardiovascular medicine and haematology
Research Interests: Biology, Biological Chemistry, DNA damage, Medicine, Biological Sciences, and 15 moreCell line, Humans, Anoxia, Protein Kinases, Mice, Animals, Phosphorylation, Proteins, Immunoprecipitation, CHEMICAL SCIENCES, DNA binding proteins, Glucocorticoids, Protein Biosynthesis, Gene Expression Regulation, and Medical and Health Sciences
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Tuberous sclerosis complex (TSC) and Peutz-Jeghers syndrome (PJS) are dominantly inherited benign tumor syndromes that share striking histopathological similarities. Here we show that LKB1, the gene mutated in PJS, acts as a tumor... more
Tuberous sclerosis complex (TSC) and Peutz-Jeghers syndrome (PJS) are dominantly inherited benign tumor syndromes that share striking histopathological similarities. Here we show that LKB1, the gene mutated in PJS, acts as a tumor suppressor by activating TSC2, the gene mutated in TSC. Like TSC2, LKB1 inhibits the phosphorylation of the key translational regulators S6K and 4EBP1. Furthermore, we show that LKB1 activates TSC2 through the AMP-dependent protein kinase (AMPK), indicating that LKB1 plays a role in cell growth regulation in response to cellular energy levels. Our results suggest that PJS and other benign tumor syndromes could be caused by dysregulation of the TSC2/mTOR pathway.