The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Yasuhiro Minami - One of the best experts on this subject based on the ideXlab platform.
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receptor tyrosine kinase ror2 mediates wnt5a induced polarized cell migration by activating c jun n terminal kinase via Actin Binding Protein filamin a
Journal of Biological Chemistry, 2008Co-Authors: Akira Nomachi, Michiru Nishita, Daisuke Inaba, Mayumi Hamasaki, Masahiro Enomoto, Yasuhiro MinamiAbstract:Abstract The receptor tyrosine kinase Ror2 has recently been shown to act as an alternative receptor or coreceptor for Wnt5a and to mediate Wnt5a-induced migration of cultured cells. However, little is known about the molecular mechanism underlying this migratory process. Here we show by wound-healing assays that Ror2 plays critical roles in Wnt5a-induced cell migration by regulating formation of lamellipodia and reorientation of microtubule-organizing center (MTOC). Wnt5a stimulation induces activation of the c-Jun N-terminal kinase JNK at the wound edge in a Ror2-dependent manner, and inhibiting JNK activity abrogates Wnt5a-induced lamellipodia formation and MTOC reorientation. Additionally, the association of Ror2 with the Actin-Binding Protein filamin A is required for Wnt5a-induced JNK activation and polarized cell migration. We further show that Wnt5a-induced JNK activation and MTOC reorientation can be suppressed by inhibiting PKCζ. Taken together, our findings indicate that Wnt5a/Ror2 activates JNK, through a process involving filamin A and PKCζ, to regulate polarized cell migration.
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receptor tyrosine kinase ror2 mediates wnt5a induced polarized cell migration by activating c jun n terminal kinase via Actin Binding Protein filamin a
Journal of Biological Chemistry, 2008Co-Authors: Akira Nomachi, Michiru Nishita, Daisuke Inaba, Mayumi Hamasaki, Masahiro Enomoto, Yasuhiro MinamiAbstract:The receptor tyrosine kinase Ror2 has recently been shown to act as an alternative receptor or coreceptor for Wnt5a and to mediate Wnt5a-induced migration of cultured cells. However, little is known about the molecular mechanism underlying this migratory process. Here we show by wound-healing assays that Ror2 plays critical roles in Wnt5a-induced cell migration by regulating formation of lamellipodia and reorientation of microtubule-organizing center (MTOC). Wnt5a stimulation induces activation of the c-Jun N-terminal kinase JNK at the wound edge in a Ror2-dependent manner, and inhibiting JNK activity abrogates Wnt5a-induced lamellipodia formation and MTOC reorientation. Additionally, the association of Ror2 with the Actin-Binding Protein filamin A is required for Wnt5a-induced JNK activation and polarized cell migration. We further show that Wnt5a-induced JNK activation and MTOC reorientation can be suppressed by inhibiting PKCzeta. Taken together, our findings indicate that Wnt5a/Ror2 activates JNK, through a process involving filamin A and PKCzeta, to regulate polarized cell migration.
Akira Nomachi - One of the best experts on this subject based on the ideXlab platform.
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receptor tyrosine kinase ror2 mediates wnt5a induced polarized cell migration by activating c jun n terminal kinase via Actin Binding Protein filamin a
Journal of Biological Chemistry, 2008Co-Authors: Akira Nomachi, Michiru Nishita, Daisuke Inaba, Mayumi Hamasaki, Masahiro Enomoto, Yasuhiro MinamiAbstract:Abstract The receptor tyrosine kinase Ror2 has recently been shown to act as an alternative receptor or coreceptor for Wnt5a and to mediate Wnt5a-induced migration of cultured cells. However, little is known about the molecular mechanism underlying this migratory process. Here we show by wound-healing assays that Ror2 plays critical roles in Wnt5a-induced cell migration by regulating formation of lamellipodia and reorientation of microtubule-organizing center (MTOC). Wnt5a stimulation induces activation of the c-Jun N-terminal kinase JNK at the wound edge in a Ror2-dependent manner, and inhibiting JNK activity abrogates Wnt5a-induced lamellipodia formation and MTOC reorientation. Additionally, the association of Ror2 with the Actin-Binding Protein filamin A is required for Wnt5a-induced JNK activation and polarized cell migration. We further show that Wnt5a-induced JNK activation and MTOC reorientation can be suppressed by inhibiting PKCζ. Taken together, our findings indicate that Wnt5a/Ror2 activates JNK, through a process involving filamin A and PKCζ, to regulate polarized cell migration.
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receptor tyrosine kinase ror2 mediates wnt5a induced polarized cell migration by activating c jun n terminal kinase via Actin Binding Protein filamin a
Journal of Biological Chemistry, 2008Co-Authors: Akira Nomachi, Michiru Nishita, Daisuke Inaba, Mayumi Hamasaki, Masahiro Enomoto, Yasuhiro MinamiAbstract:The receptor tyrosine kinase Ror2 has recently been shown to act as an alternative receptor or coreceptor for Wnt5a and to mediate Wnt5a-induced migration of cultured cells. However, little is known about the molecular mechanism underlying this migratory process. Here we show by wound-healing assays that Ror2 plays critical roles in Wnt5a-induced cell migration by regulating formation of lamellipodia and reorientation of microtubule-organizing center (MTOC). Wnt5a stimulation induces activation of the c-Jun N-terminal kinase JNK at the wound edge in a Ror2-dependent manner, and inhibiting JNK activity abrogates Wnt5a-induced lamellipodia formation and MTOC reorientation. Additionally, the association of Ror2 with the Actin-Binding Protein filamin A is required for Wnt5a-induced JNK activation and polarized cell migration. We further show that Wnt5a-induced JNK activation and MTOC reorientation can be suppressed by inhibiting PKCzeta. Taken together, our findings indicate that Wnt5a/Ror2 activates JNK, through a process involving filamin A and PKCzeta, to regulate polarized cell migration.
David G Drubin - One of the best experts on this subject based on the ideXlab platform.
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direct regulation of arp2 3 complex activity and function by the Actin Binding Protein coronin
Journal of Cell Biology, 2002Co-Authors: Christine Humphries, David G Drubin, Heath I Balcer, Jessica L Dagostino, Barbara Winsor, Georjana Barnes, Brenda J Andrews, Bruce L GoodeAbstract:Mechanisms for activating the Actin-related Protein 2/3 (Arp2/3) complex have been the focus of many recent studies. Here, we identify a novel mode of Arp2/3 complex regulation mediated by the highly conserved Actin Binding Protein coronin. Yeast coronin (Crn1) physically associates with the Arp2/3 complex and inhibits WA- and Abp1-activated Actin nucleation in vitro. The inhibition occurs specifically in the absence of preformed Actin filaments, suggesting that Crn1 may restrict Arp2/3 complex activity to the sides of filaments. The inhibitory activity of Crn1 resides in its coiled coil domain. Localization of Crn1 to Actin patches in vivo and association of Crn1 with the Arp2/3 complex also require its coiled coil domain. Genetic studies provide in vivo evidence for these interactions and activities. Overexpression of CRN1 causes growth arrest and redistribution of Arp2 and Crn1p into aberrant Actin loops. These defects are suppressed by deletion of the Crn1 coiled coil domain and by arc35-26, an allele of the p35 subunit of the Arp2/3 complex. Further in vivo evidence that coronin regulates the Arp2/3 complex comes from the observation that crn1 and arp2 mutants display an allele-specific synthetic interaction. This work identifies a new form of regulation of the Arp2/3 complex and an important cellular function for coronin.
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unusual Binding properties of the sh3 domain of the yeast Actin Binding Protein abp1 structural and functional analysis
Journal of Biological Chemistry, 2002Co-Authors: Barbara Fazi, David G Drubin, Jamie M T V Cope, Alice Douangamath, Silvia Ferracuti, Katja Schirwitz, Adriana Zucconi, Matthias Wilmanns, Gianni Cesareni, Luisa CastagnoliAbstract:Abp1p is an Actin-Binding Protein that plays a central role in the organization of Saccharomyces cerevisiae Actin cytoskeleton. By a combination of two-hybrid and phage-display approaches, we have identified six new ligands of the Abp1-SH3 domain. None of these SH3-mediated novel interactions was detected in recent all genome high throughput Protein interaction projects. Here we show that the SH3-mediated association of Abp1p with the Ser/Thr kinases Prk1p and Ark1p is essential for their localization to Actin cortical patches. The Abp1-SH3 domain has a rather unusual Binding specificity, because its target peptides contain the tetrapentapeptide +XXXPXXPX+PXXL with positive charges flanking the polyproline core on both sides. Here we present the structure of the Abp1-SH3 domain solved at 1.3-A resolution. The peptide-Binding pockets in the SH3 domain are flanked by two acidic residues that are uncommon at those positions in the SH3 domain family. We have shown by site-directed mutagenesis that one of these negatively charged side chains may be the key determinant for the preference for non-classical ligands.
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the Actin Binding Protein hip1r associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro
Journal of Cell Biology, 2001Co-Authors: Asa E Y Engqvistgoldstein, Robin A Warren, Michael M Kessels, James H Keen, John E Heuser, David G DrubinAbstract:Huntingtin-interActing Protein 1 related (Hip1R) is a novel component of clathrin-coated pits and vesicles and is a mammalian homologue of Sla2p, an Actin-Binding Protein important for both Actin organization and endocytosis in yeast. Here, we demonstrate that Hip1R binds via its putative central coiled-coil domain to clathrin, and provide evidence that Hip1R and clathrin are associated in vivo at sites of endocytosis. First, real-time analysis of Hip1R–YFP and DsRed–clathrin light chain (LC) in live cells revealed that these Proteins show almost identical temporal and spatial regulation at the cell cortex. Second, at the ultrastructure level, immunogold labeling of ‘unroofed’ cells showed that Hip1R localizes to clathrin-coated pits. Third, overexpression of Hip1R affected the subcellular distribution of clathrin LC. Consistent with a functional role for Hip1R in endocytosis, we also demonstrated that it promotes clathrin cage assembly in vitro. Finally, we showed that Hip1R is a rod-shaped apparent dimer with globular heads at either end, and that it can assemble clathrin-coated vesicles and F-Actin into higher order structures. In total, Hip1R's properties suggest an early endocytic function at the interface between clathrin, F-Actin, and lipids.
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mammalian abp1 a signal responsive f Actin Binding Protein links the Actin cytoskeleton to endocytosis via the gtpase dynamin
Journal of Cell Biology, 2001Co-Authors: Michael M Kessels, David G Drubin, Asa E Y Engqvistgoldstein, Britta QualmannAbstract:The Actin cytoskeleton has been implicated in endocytosis, yet few molecular links to the endocytic machinery have been established. Here we show that the mammalian F-Actin–Binding Protein Abp1 (SH3P7/HIP-55) can functionally link the Actin cytoskeleton to dynamin, a GTPase that functions in endocytosis. Abp1 binds directly to dynamin in vitro through its SH3 domain. Coimmunoprecipitation and colocalization studies demonstrated the in vivo relevance of this interaction. In neurons, mammalian Abp1 and dynamin colocalized at Actin-rich sites proximal to the cell body during synaptogenesis. In fibroblasts, mAbp1 appeared at dynamin-rich sites of endocytosis upon growth factor stimulation. To test whether Abp1 functions in endocytosis, we overexpressed several Abp1 constructs in Cos-7 cells and assayed receptor-mediated endocytosis. While overexpression of Abp1's Actin-Binding modules did not interfere with endocytosis, overexpression of the SH3 domain led to a potent block of transferrin uptake. This implicates the Abp1/dynamin interaction in endocytic function. The endocytosis block was rescued by cooverexpression of dynamin. Since the addition of the Actin-Binding modules of Abp1 to the SH3 domain construct also fully restored endocytosis, Abp1 may support endocytosis by combining its SH3 domain interactions with cytoskeletal functions in response to signaling cascades converging on this linker Protein.
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an Actin Binding Protein of the sla2 huntingtin interActing Protein 1 family is a novel component of clathrin coated pits and vesicles
Journal of Cell Biology, 1999Co-Authors: Asa E Y Engqvistgoldstein, Michael M Kessels, Vikramjit Chopra, Michael R Hayden, David G DrubinAbstract:The Actin cytoskeleton has been implicated in endocytosis, yet few molecules that link these systems have been identified. Here, we have cloned and characterized mHip1R, a Protein that is closely related to huntingtin interActing Protein 1 (Hip1). These two Proteins are mammalian homologues of Sla2p, an Actin Binding Protein important for Actin organization and endocytosis in yeast. Sequence alignments and secondary structure predictions verified that mHip1R belongs to the Sla2 Protein family. Thus, mHip1R contains an NH2-terminal domain homologous to that implicated in Sla2p's endocytic function, three predicted coiled–coils, a leucine zipper, and a talin-like Actin-Binding domain at the COOH terminus. The talin-like domain of mHip1R binds to F-Actin in vitro and colocalizes with F-Actin in vivo, indicating that this activity has been conserved from yeast to mammals. mHip1R shows a punctate immunolocalization and is enriched at the cell cortex and in the perinuclear region. We concluded that the cortical localization represents endocytic compartments, because mHip1R colocalizes with clathrin, AP-2, and endocytosed transferrin, and because mHip1R fractionates biochemically with clathrin-coated vesicles. Time-lapse video microscopy of mHip1R–green fluorescence Protein (GFP) revealed a blinking behavior similar to that reported for GFP-clathrin, and an Actin-dependent inward movement of punctate structures from the cell periphery. These data show that mHip1R is a component of clathrin-coated pits and vesicles and suggest that it might link the endocytic machinery to the Actin cytoskeleton.
Mayumi Hamasaki - One of the best experts on this subject based on the ideXlab platform.
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receptor tyrosine kinase ror2 mediates wnt5a induced polarized cell migration by activating c jun n terminal kinase via Actin Binding Protein filamin a
Journal of Biological Chemistry, 2008Co-Authors: Akira Nomachi, Michiru Nishita, Daisuke Inaba, Mayumi Hamasaki, Masahiro Enomoto, Yasuhiro MinamiAbstract:Abstract The receptor tyrosine kinase Ror2 has recently been shown to act as an alternative receptor or coreceptor for Wnt5a and to mediate Wnt5a-induced migration of cultured cells. However, little is known about the molecular mechanism underlying this migratory process. Here we show by wound-healing assays that Ror2 plays critical roles in Wnt5a-induced cell migration by regulating formation of lamellipodia and reorientation of microtubule-organizing center (MTOC). Wnt5a stimulation induces activation of the c-Jun N-terminal kinase JNK at the wound edge in a Ror2-dependent manner, and inhibiting JNK activity abrogates Wnt5a-induced lamellipodia formation and MTOC reorientation. Additionally, the association of Ror2 with the Actin-Binding Protein filamin A is required for Wnt5a-induced JNK activation and polarized cell migration. We further show that Wnt5a-induced JNK activation and MTOC reorientation can be suppressed by inhibiting PKCζ. Taken together, our findings indicate that Wnt5a/Ror2 activates JNK, through a process involving filamin A and PKCζ, to regulate polarized cell migration.
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receptor tyrosine kinase ror2 mediates wnt5a induced polarized cell migration by activating c jun n terminal kinase via Actin Binding Protein filamin a
Journal of Biological Chemistry, 2008Co-Authors: Akira Nomachi, Michiru Nishita, Daisuke Inaba, Mayumi Hamasaki, Masahiro Enomoto, Yasuhiro MinamiAbstract:The receptor tyrosine kinase Ror2 has recently been shown to act as an alternative receptor or coreceptor for Wnt5a and to mediate Wnt5a-induced migration of cultured cells. However, little is known about the molecular mechanism underlying this migratory process. Here we show by wound-healing assays that Ror2 plays critical roles in Wnt5a-induced cell migration by regulating formation of lamellipodia and reorientation of microtubule-organizing center (MTOC). Wnt5a stimulation induces activation of the c-Jun N-terminal kinase JNK at the wound edge in a Ror2-dependent manner, and inhibiting JNK activity abrogates Wnt5a-induced lamellipodia formation and MTOC reorientation. Additionally, the association of Ror2 with the Actin-Binding Protein filamin A is required for Wnt5a-induced JNK activation and polarized cell migration. We further show that Wnt5a-induced JNK activation and MTOC reorientation can be suppressed by inhibiting PKCzeta. Taken together, our findings indicate that Wnt5a/Ror2 activates JNK, through a process involving filamin A and PKCzeta, to regulate polarized cell migration.
Michiru Nishita - One of the best experts on this subject based on the ideXlab platform.
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receptor tyrosine kinase ror2 mediates wnt5a induced polarized cell migration by activating c jun n terminal kinase via Actin Binding Protein filamin a
Journal of Biological Chemistry, 2008Co-Authors: Akira Nomachi, Michiru Nishita, Daisuke Inaba, Mayumi Hamasaki, Masahiro Enomoto, Yasuhiro MinamiAbstract:Abstract The receptor tyrosine kinase Ror2 has recently been shown to act as an alternative receptor or coreceptor for Wnt5a and to mediate Wnt5a-induced migration of cultured cells. However, little is known about the molecular mechanism underlying this migratory process. Here we show by wound-healing assays that Ror2 plays critical roles in Wnt5a-induced cell migration by regulating formation of lamellipodia and reorientation of microtubule-organizing center (MTOC). Wnt5a stimulation induces activation of the c-Jun N-terminal kinase JNK at the wound edge in a Ror2-dependent manner, and inhibiting JNK activity abrogates Wnt5a-induced lamellipodia formation and MTOC reorientation. Additionally, the association of Ror2 with the Actin-Binding Protein filamin A is required for Wnt5a-induced JNK activation and polarized cell migration. We further show that Wnt5a-induced JNK activation and MTOC reorientation can be suppressed by inhibiting PKCζ. Taken together, our findings indicate that Wnt5a/Ror2 activates JNK, through a process involving filamin A and PKCζ, to regulate polarized cell migration.
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receptor tyrosine kinase ror2 mediates wnt5a induced polarized cell migration by activating c jun n terminal kinase via Actin Binding Protein filamin a
Journal of Biological Chemistry, 2008Co-Authors: Akira Nomachi, Michiru Nishita, Daisuke Inaba, Mayumi Hamasaki, Masahiro Enomoto, Yasuhiro MinamiAbstract:The receptor tyrosine kinase Ror2 has recently been shown to act as an alternative receptor or coreceptor for Wnt5a and to mediate Wnt5a-induced migration of cultured cells. However, little is known about the molecular mechanism underlying this migratory process. Here we show by wound-healing assays that Ror2 plays critical roles in Wnt5a-induced cell migration by regulating formation of lamellipodia and reorientation of microtubule-organizing center (MTOC). Wnt5a stimulation induces activation of the c-Jun N-terminal kinase JNK at the wound edge in a Ror2-dependent manner, and inhibiting JNK activity abrogates Wnt5a-induced lamellipodia formation and MTOC reorientation. Additionally, the association of Ror2 with the Actin-Binding Protein filamin A is required for Wnt5a-induced JNK activation and polarized cell migration. We further show that Wnt5a-induced JNK activation and MTOC reorientation can be suppressed by inhibiting PKCzeta. Taken together, our findings indicate that Wnt5a/Ror2 activates JNK, through a process involving filamin A and PKCzeta, to regulate polarized cell migration.
