TBCK Influences Cell Proliferation, Cell Size and mTOR Signaling Pathway

Mammalian target of rapamycin (mTOR) is a central regulator for both cell proliferation and cell growth; however, little is known about the regulation of mTOR expression at the transcriptional level. Here, we provide evidences that a conserved human protein TBCK (TBC1 domain containing kinase) is involved in the regulation of mTOR signaling pathway. Depletion of TBCK significantly inhibits cell proliferation, reduces cell size, and disrupts the organization of actin, but not microtubule. Knockdown of TBCK induces a significant decrease in the protein levels of components of mTOR complex (mTORC), and suppresses the activity of mTOR signaling, but not MAPK or PDK1/Akt pathway. Further results show that TBCK influences the expression of mTORC components at the transcriptional level. Thus, these data suggest that TBCK may play an important role in cell proliferation, cell growth and actin organization possibly by modulating mTOR pathway.

[1]  R. Roth,et al.  Akt promotes increased mammalian cell size by stimulating protein synthesis and inhibiting protein degradation. , 2003, American journal of physiology. Endocrinology and metabolism.

[2]  Jacob D. Jaffe,et al.  mSin1 Is Necessary for Akt/PKB Phosphorylation, and Its Isoforms Define Three Distinct mTORC2s , 2006, Current Biology.

[3]  T. Hunter,et al.  The eukaryotic protein kinase superfamily: kinase (catalytic) domain structure and classification 1 , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[4]  E. Birney,et al.  Patterns of somatic mutation in human cancer genomes , 2007, Nature.

[5]  P. Finan,et al.  TBC1D7 is a third subunit of the TSC1-TSC2 complex upstream of mTORC1. , 2012, Molecular cell.

[6]  Kassi Miller,et al.  Exercise modulates microRNAs that affect the PTEN/mTOR pathway in rats after spinal cord injury , 2012, Experimental Neurology.

[7]  Ernesto Picardi,et al.  UTRdb and UTRsite (RELEASE 2010): a collection of sequences and regulatory motifs of the untranslated regions of eukaryotic mRNAs , 2009, Nucleic Acids Res..

[8]  D. Sabatini,et al.  mTOR Signaling in Growth Control and Disease , 2012, Cell.

[9]  G. Barton,et al.  Emerging roles of pseudokinases. , 2006, Trends in cell biology.

[10]  D. Guertin,et al.  Rictor, a Novel Binding Partner of mTOR, Defines a Rapamycin-Insensitive and Raptor-Independent Pathway that Regulates the Cytoskeleton , 2004, Current Biology.

[11]  J. Qin,et al.  SIN1/MIP1 Maintains rictor-mTOR Complex Integrity and Regulates Akt Phosphorylation and Substrate Specificity , 2006, Cell.

[12]  Paul Tempst,et al.  GbetaL, a positive regulator of the rapamycin-sensitive pathway required for the nutrient-sensitive interaction between raptor and mTOR. , 2003, Molecular cell.

[13]  Ling Tian,et al.  MicroRNA‐7 inhibits tumor growth and metastasis by targeting the phosphoinositide 3‐kinase/Akt pathway in hepatocellular carcinoma , 2012, Hepatology.

[14]  A. Nakashima,et al.  Identification of TBC7 having TBC domain as a novel binding protein to TSC1-TSC2 complex. , 2007, Biochemical and biophysical research communications.

[15]  Timothy J. Griffin,et al.  Insulin signalling to mTOR mediated by the Akt/PKB substrate PRAS40 , 2007, Nature Cell Biology.

[16]  H. Okano,et al.  Musashi: a translational regulator of cell fate. , 2002, Journal of cell science.

[17]  J. Avruch,et al.  Raptor, a Binding Partner of Target of Rapamycin (TOR), Mediates TOR Action , 2002, Cell.

[18]  P. Bucher,et al.  A model of Cdc25 phosphatase catalytic domain and Cdk-interaction surface based on the presence of a rhodanese homology domain. , 1998, Journal of molecular biology.

[19]  T. Hunter,et al.  The eukaryotic protein kinase superfamily: kinase (catalytic) domain structure and classification 1 , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[20]  N. Sonenberg,et al.  Upstream and downstream of mTOR. , 2004, Genes & development.

[21]  D. Alessi,et al.  Protor-1 is required for efficient mTORC2-mediated activation of SGK1 in the kidney. , 2011, The Biochemical journal.

[22]  G. Wakabayashi,et al.  Downregulation of miR-144 is associated with colorectal cancer progression via activation of mTOR signaling pathway. , 2012, Carcinogenesis.

[23]  H. Stenmark Rab GTPases as coordinators of vesicle traffic , 2009, Nature Reviews Molecular Cell Biology.

[24]  E. Morii,et al.  MicroRNA-mediated downregulation of mTOR/FGFR3 controls tumor growth induced by Src-related oncogenic pathways , 2011, Oncogene.

[25]  R. Loewith,et al.  Mammalian TOR complex 2 controls the actin cytoskeleton and is rapamycin insensitive , 2004, Nature Cell Biology.

[26]  Peer Bork,et al.  The rhodanese/Cdc25 phosphatase superfamily , 2002, EMBO reports.

[27]  Yusuke Nakamura,et al.  Activation of an oncogenic TBC1D7 (TBC1 domain family, member 7) protein in pulmonary carcinogenesis , 2010, Genes, chromosomes & cancer.

[28]  D. Lambright,et al.  TBC-domain GAPs for Rab GTPases accelerate GTP hydrolysis by a dual-finger mechanism , 2006, Nature.

[29]  M. Fukuda,et al.  Screening for target Rabs of TBC (Tre‐2/Bub2/Cdc16) domain‐containing proteins based on their Rab‐binding activity , 2006, Genes to cells : devoted to molecular & cellular mechanisms.

[30]  P. Zhang,et al.  MicroRNA-99a Inhibits Hepatocellular Carcinoma Growth and Correlates with Prognosis of Patients with Hepatocellular Carcinoma* , 2011, The Journal of Biological Chemistry.

[31]  D. Lambright,et al.  Rab GEFs and GAPs. , 2010, Current opinion in cell biology.

[32]  M. Hall,et al.  TOR Signaling in Growth and Metabolism , 2006, Cell.

[33]  K. Mikoshiba,et al.  Mouse-Musashi-1, a neural RNA-binding protein highly enriched in the mammalian CNS stem cell. , 1996, Developmental biology.

[34]  K. Guan,et al.  Amino acid signaling in TOR activation. , 2011, Annual review of biochemistry.

[35]  Stefan Knapp,et al.  Structure of the Pseudokinase VRK3 Reveals a Degraded Catalytic Site, a Highly Conserved Kinase Fold, and a Putative Regulatory Binding Site , 2009, Structure.

[36]  C. Croce,et al.  MiR-199a-3p regulates mTOR and c-Met to influence the doxorubicin sensitivity of human hepatocarcinoma cells. , 2010, Cancer research.

[37]  D. Alessi,et al.  Identification of Protor as a novel Rictor-binding component of mTOR complex-2. , 2007, The Biochemical journal.

[38]  Obi L. Griffith,et al.  cisRED: a database system for genome-scale computational discovery of regulatory elements , 2005, Nucleic Acids Res..

[39]  E. Birney,et al.  Patterns of somatic mutation in human cancer genomes , 2007, Nature.

[40]  Tobias Schmelzle,et al.  TOR, a Central Controller of Cell Growth , 2000, Cell.

[41]  P. Novick,et al.  Role of Rab GTPases in membrane traffic and cell physiology. , 2011, Physiological reviews.

[42]  Adiel Cohen,et al.  PRAS40 and PRR5-Like Protein Are New mTOR Interactors that Regulate Apoptosis , 2007, PloS one.

[43]  R. Roth,et al.  PRAS40 Regulates mTORC1 Kinase Activity by Functioning as a Direct Inhibitor of Substrate Binding* , 2007, Journal of Biological Chemistry.

[44]  T. Hunter,et al.  The Protein Kinase Complement of the Human Genome , 2002, Science.

[45]  D. Sabatini,et al.  mTOR Interacts with Raptor to Form a Nutrient-Sensitive Complex that Signals to the Cell Growth Machinery , 2002, Cell.

[46]  Tohru Natsume,et al.  Tti1 and Tel2 Are Critical Factors in Mammalian Target of Rapamycin Complex Assembly* , 2010, The Journal of Biological Chemistry.

[47]  D. Sabatini,et al.  DEPTOR Is an mTOR Inhibitor Frequently Overexpressed in Multiple Myeloma Cells and Required for Their Survival , 2009, Cell.

[48]  Li Lin,et al.  Identification of miRNomes in human liver and hepatocellular carcinoma reveals miR-199a/b-3p as therapeutic target for hepatocellular carcinoma. , 2011, Cancer cell.

[49]  M. Hall,et al.  Target of Rapamycin (TOR) in Nutrient Signaling and Growth Control , 2011, Genetics.

[50]  M. Srougi,et al.  MicroRNA-100 expression is independently related to biochemical recurrence of prostate cancer. , 2011, The Journal of urology.

[51]  B. Manning,et al.  The TSC1-TSC2 complex: a molecular switchboard controlling cell growth. , 2008, The Biochemical journal.

[52]  Paul Lasko,et al.  Translational control in cellular and developmental processes , 2012, Nature Reviews Genetics.

[53]  S. Carr,et al.  PRAS40 is an insulin-regulated inhibitor of the mTORC1 protein kinase. , 2007, Molecular cell.

[54]  M. Ahmadian,et al.  Illuminating the functional and structural repertoire of human TBC/RABGAPs , 2012, Nature Reviews Molecular Cell Biology.