The Ankyrin Repeat-containing Adaptor Protein Tvl-1 Is a Novel Substrate and Regulator of Raf-1*

Tvl-1 is a 269-amino acid ankyrin repeat protein expressed primarily in thymus, lung, and testes that was identified by screening a murine T-cell two-hybrid cDNA library for proteins that associate with the serine-threonine kinase Raf-1. The interaction of Tvl-1 with Raf-1 was confirmed by co-immunoprecipitation of the two proteins from COS-1 cells transiently transfected with Tvl-1 and Raf-1 expression constructs as well as by co-immunoprecipitation of the endogenous proteins from CV-1 and NB2 cells. Tvl-1 interacts with Raf-1 via its carboxyl-terminal ankyrin repeat domain. The same domain also mediates Tvl-1 homodimerization. Tvl-1 was detected by immunofluorescence in both the cytoplasm and the nucleus suggesting that in addition to Raf-1 it may also interact with nuclear proteins. Activated Raf-1 phosphorylates Tvl-1 both in vitro andin vivo. In baculovirus-infected Sf9 insect cells, Tvl-1 potentiates the activation of Raf-1 by Src and Ras while in COS-1 cells it potentiates the activation of Raf-1 by EGF. These data suggest that Tvl-1 is both a target as well as a regulator of Raf-1. The human homologue of Tvl-1 maps to chromosome 19p12, upstream ofMEF2B with the two genes in a head to head arrangement.

[1]  B. Cochran,et al.  p50cdc37 Acting in Concert with Hsp90 Is Required for Raf-1 Function , 1999, Molecular and Cellular Biology.

[2]  R. Aebersold,et al.  A gene encoding a novel RFX-associated transactivator is mutated in the majority of MHC class II deficiency patients , 1998, Nature Genetics.

[3]  S. Artavanis-Tsakonas,et al.  Suppressor of Hairless-independent events in Notch signaling imply novel pathway elements. , 1997, Development.

[4]  F. McCormick,et al.  Activation of c‐Raf‐1 by Ras and Src through different mechanisms: activation in vivo and in vitro , 1997, The EMBO journal.

[5]  D. Morrison,et al.  The complexity of Raf-1 regulation. , 1997, Current opinion in cell biology.

[6]  John Calvin Reed,et al.  Bcl-2 Targets the Protein Kinase Raf-1 to Mitochondria , 1996, Cell.

[7]  P. Pitha,et al.  Binding of human immunodeficiency virus type 1 to CD4 induces association of Lck and Raf-1 and activates Raf-1 by a Ras-independent pathway , 1996, Molecular and cellular biology.

[8]  M. Farrar,et al.  Activation of the Raf-1 kinase cascade by coumermycin-induced dimerization , 1996, Nature.

[9]  P. J. Belshaw,et al.  Oligomerization activates c-Raf-1 through a Ras-dependent mechanism , 1996, Nature.

[10]  J. Harper,et al.  Mammalian p50Cdc37 is a protein kinase-targeting subunit of Hsp90 that binds and stabilizes Cdk4. , 1996, Genes & development.

[11]  D. Durden,et al.  A Role for Shc, Grb2, and Raf-1 in FcγRI Signal Relay* , 1996, The Journal of Biological Chemistry.

[12]  R. Bell,et al.  Raf-1 Kinase Possesses Distinct Binding Domains for Phosphatidylserine and Phosphatidic Acid , 1996, The Journal of Biological Chemistry.

[13]  P. Dent,et al.  Inactivation of Raf-1 by a Protein-tyrosine Phosphatase Stimulated by GTP and Reconstituted by G Subunits (*) , 1996, The Journal of Biological Chemistry.

[14]  M. White,et al.  Ras Interaction with Two Distinct Binding Domains in Raf-1 5 Be Required for Ras Transformation (*) , 1996, The Journal of Biological Chemistry.

[15]  Mikhail V. Blagosklonny,et al.  Disruption of the Raf-1-Hsp90 Molecular Complex Results in Destabilization of Raf-1 and Loss of Raf-1-Ras Association (*) , 1995, The Journal of Biological Chemistry.

[16]  M. Siciliano,et al.  Regional chromosomal assignments for four members of the MADS domain transcription enhancer factor 2 (MEF2) gene family to human chromosomes 15q26, 19p12, 5q14, and 1q12-q23. , 1995, Genomics.

[17]  X. F. Zhang,et al.  Identification of the 14.3.3 ζ Domains Important for Self-association and Raf Binding (*) , 1995, The Journal of Biological Chemistry.

[18]  D. Morrison,et al.  Regulation of Raf-1 and Raf-1 mutants by Ras-dependent and Ras-independent mechanisms in vitro , 1995, Molecular and cellular biology.

[19]  C. Marshall,et al.  Ras recruits Raf‐1 to the plasma membrane for activation by tyrosine phosphorylation. , 1995, The EMBO journal.

[20]  D. Beach,et al.  Raf1 interaction with Cdc25 phosphatase ties mitogenic signal transduction to cell cycle activation. , 1995, Genes & development.

[21]  C. Der,et al.  Two Distinct Raf Domains Mediate Interaction with Ras (*) , 1995, The Journal of Biological Chemistry.

[22]  R. Hipskind,et al.  Ras-dependent and -independent pathways target the mitogen-activated protein kinase network in macrophages , 1995, Molecular and cellular biology.

[23]  R. Brent,et al.  Interaction mating reveals binary and ternary connections between Drosophila cell cycle regulators. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[24]  R. Bell,et al.  Identification of discrete segments of human Raf-1 kinase critical for high affinity binding to Ha-Ras. , 1994, The Journal of biological chemistry.

[25]  W. Fantl,et al.  Activation of Raf-1 by 14-3-3 proteins , 1994, Nature.

[26]  P. Tsichlis,et al.  Tpl-2 acts in concert with Ras and Raf-1 to activate mitogen-activated protein kinase. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[27]  W. Kolch,et al.  Mechanism of inhibition of Raf-1 by protein kinase A , 1994, Molecular and cellular biology.

[28]  K. Irie,et al.  Stimulatory effects of yeast and mammalian 14-3-3 proteins on the Raf protein kinase. , 1994, Science.

[29]  F. McCormick,et al.  Binding of 14-3-3 proteins to the protein kinase Raf and effects on its activation. , 1994, Science.

[30]  X. F. Zhang,et al.  Critical binding and regulatory interactions between Ras and Raf occur through a small, stable N-terminal domain of Raf and specific Ras effector residues , 1994, Molecular and cellular biology.

[31]  X. F. Zhang,et al.  Raf meets Ras: completing the framework of a signal transduction pathway. , 1994, Trends in biochemical sciences.

[32]  J. Hancock,et al.  Activation of Raf as a result of recruitment to the plasma membrane. , 1994, Science.

[33]  Sally J. Leevers,et al.  Requirement for Ras in Raf activation is overcome by targeting Raf to the plasma membrane , 1994, Nature.

[34]  R. Davis,et al.  The native structure of the activated Raf protein kinase is a membrane-bound multi-subunit complex. , 1994, The Journal of biological chemistry.

[35]  C. Zheng,et al.  Activation of MEK family kinases requires phosphorylation of two conserved Ser/Thr residues. , 1994, The EMBO journal.

[36]  V. Bennett,et al.  440-kD ankyrinB: structure of the major developmentally regulated domain and selective localization in unmyelinated axons , 1993, The Journal of cell biology.

[37]  Roger Brent,et al.  C dil, a Human Gl and S Phase Protein Phosphatase That Associates with Cdk2 , 2003 .

[38]  F. McCormick,et al.  Reconstitution of the Raf-1—MEK—ERK Signal Transduction Pathway In Vitro , 1993, Molecular and cellular biology.

[39]  D. Morrison,et al.  Critical tyrosine residues regulate the enzymatic and biological activity of Raf-1 kinase , 1993, Molecular and cellular biology.

[40]  G. Johnson,et al.  Activation of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase by G protein and tyrosine kinase oncoproteins. , 1993, The Journal of biological chemistry.

[41]  D. Morrison,et al.  Identification of the major phosphorylation sites of the Raf-1 kinase. , 1993, The Journal of biological chemistry.

[42]  J. Avruch,et al.  Mitogen regulation of c-Raf-1 protein kinase activity toward mitogen-activated protein kinase-kinase. , 1993, The Journal of biological chemistry.

[43]  S. Elledge,et al.  Normal and oncogenic p21ras proteins bind to the amino-terminal regulatory domain of c-Raf-1 , 1993, Nature.

[44]  P. Warne,et al.  Direct interaction of Ras and the amino-terminal region of Raf-1 in vitro , 1993, Nature.

[45]  Jonathan A. Cooper,et al.  Mammalian Ras interacts directly with the serine/threonine kinase raf , 1993, Cell.

[46]  Walter Kolch,et al.  Protein kinase Cα activates RAF-1 by direct phosphorylation , 1993, Nature.

[47]  P. Cohen,et al.  Activation of the MAP kinase pathway by the protein kinase raf , 1992, Cell.

[48]  T. Haystead,et al.  Activation of mitogen-activated protein kinase kinase by v-Raf in NIH 3T3 cells and in vitro. , 1992, Science.

[49]  B. Nadal-Ginard,et al.  Human myocyte-specific enhancer factor 2 comprises a group of tissue-restricted MADS box transcription factors. , 1992, Genes & development.

[50]  David L. Brautigan,et al.  Raf-1 activates MAP kinase-kinase , 1992, Nature.

[51]  V. Bennett,et al.  The ANK repeat: a ubiquitous motif involved in macromolecular recognition. , 1992, Trends in cell biology.

[52]  T. Roberts,et al.  Both p21ras and pp60v-src are required, but neither alone is sufficient, to activate the Raf-1 kinase. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[53]  Rapp Ur.  Role of Raf-1 serine/threonine protein kinase in growth factor signal transduction. , 1991 .

[54]  H. Jörnvall,et al.  Cloning, structure, and expression of the mitochondrial cytochrome P-450 sterol 26-hydroxylase, a bile acid biosynthetic enzyme. , 1989, The Journal of biological chemistry.

[55]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[56]  R. Brent,et al.  A eukaryotic transcriptional activator bearing the DNA specificity of a prokaryotic repressor , 1985, Cell.

[57]  E. Golemis,et al.  Alternative yeast two-hybrid systems. The interaction trap and interaction mating. , 1997, Methods in molecular biology.

[58]  D. Morrison.  Activation of Raf-1 by Ras in intact cells. , 1995, Methods in enzymology.

[59]  T. Hunter,et al.  Phosphopeptide mapping and phosphoamino acid analysis by two-dimensional separation on thin-layer cellulose plates. , 1991, Methods in enzymology.