65 results on '"Hiroshi Hanafusa"'
Search Results
2. The C. elegans BRCA2-ALP/Enigma Complex Regulates Axon Regeneration via a Rho GTPase-ROCK-MLC Phosphorylation Pathway
- Author
-
Tatsuhiro Shimizu, Strahil Iv. Pastuhov, Hiroshi Hanafusa, Kunihiro Matsumoto, and Naoki Hisamoto
- Subjects
Biology (General) ,QH301-705.5 - Abstract
Summary: The ability of specific neurons to regenerate their axons after injury is governed by cell-intrinsic regeneration pathways. However, the mechanisms regulating axon regeneration are not well understood. Here, we identify the brc-2 gene encoding a homolog of the mammalian BRCA2 tumor suppressor as a regulator of axon regeneration in Caenorhabditis elegans motor neurons. We show that the RHO-1/Rho GTPase-LET-502/ROCK (Rho-associated coiled-coil kinase)-regulatory non-muscle myosin light-chain (MLC-4/MLC) phosphorylation signaling pathway regulates axon regeneration. BRC-2 functions between RHO-1 and LET-502, suggesting that BRC-2 is required for the activation of LET-502 by RHO-1-GTP. We also find that one component that interacts with BRC-2, the ALP (α-actinin-associated LIM protein)/Enigma protein ALP-1, is required for regeneration and acts between LET-502 and MLC-4 phosphorylation. Furthermore, we demonstrate that ALP-1 associates with LET-502 and MLC-4. Thus, ALP-1 serves as a platform to activate MLC-4 phosphorylation mediated by the RHO-1–LET-502 signaling pathway. : Shimizu et al. demonstrate that a BRCA2 homolog BRC-2 and its binding partner, the ALP/Enigma protein ALP-1, regulate axon regeneration via the Rho-ROCK-MLC signaling pathway in C. elegans. ALP-1 serves as a platform to activate MLC phosphorylation, which promotes growth cone formation of severed axons. Keywords: axon regeneration, BRCA2, Rho kinase, C. elegans
- Published
- 2018
- Full Text
- View/download PDF
3. Virtual grid for renewable energy society.
- Author
-
Haruhisa Ichikawa, Ashir Ahmed, Hiroshi Hanafusa, Shinji Yokogawa, Yuusuke Kawakita, Kenji Sawada, Hirohide Mikami, and Noriaki Yoshikawa
- Published
- 2015
- Full Text
- View/download PDF
4. LRRK1 functions as a scaffold for PTP1B-mediated EGFR sorting into ILVs at the ER–endosome contact site
- Author
-
Hiroshi Hanafusa, Keitaro Fujita, Misa Kamio, Shiori Iida, Yasushi Tamura, Naoki Hisamoto, and Kunihiro Matsumoto
- Subjects
Cell Biology - Abstract
Proper control of epidermal growth factor receptor (EGFR) signaling is important for maintaining cellular homeostasis. Given that EGFR signaling occurs at the plasma membrane and endosomes following internalization, endosomal trafficking of EGFR spatiotemporally regulates EGFR signaling. In this process, leucine-rich repeat kinase 1 (LRRK1) has multiple roles in kinase activity-dependent transport of EGFR-containing endosomes and kinase-independent sorting of EGFR into the intraluminal vesicles (ILVs) of multivesicular bodies. Active, phosphorylated EGFR inactivates the LRRK1 kinase activity by phosphorylating Y944. In this study, we demonstrate that LRRK1 facilitates EGFR dephosphorylation by PTP1B (also known as PTPN1), an endoplasmic reticulum (ER)-localized protein tyrosine phosphatase, at the ER–endosome contact site, after which EGFR is sorted into the ILVs of endosomes. LRRK1 is required for the PTP1B–EGFR interaction in response to EGF stimulation, resulting in the downregulation of EGFR signaling. Furthermore, PTP1B activates LRRK1 by dephosphorylating pY944 on the contact site, which promotes the transport of EGFR-containing endosomes to the perinuclear region. These findings provide evidence that the ER–endosome contact site functions as a hub for LRRK1-dependent signaling that regulates EGFR trafficking.
- Published
- 2023
5. LRRK1-mediated NDEL1 phosphorylation promotes cilia disassembly via dynein-2-driven retrograde intraflagellar transport
- Author
-
Hiroshi Hanafusa, Shin Kedashiro, Mako Gotoh, Ko-hei Saitoh, Hironori Inaba, Tomoki Nishioka, Kozo Kaibuchi, Masaki Inagaki, Naoki Hisamoto, and Kunihiro Matsumoto
- Subjects
Organelles ,Dyneins ,Biological Transport ,Cilia ,Cell Biology ,Phosphorylation - Abstract
Primary cilia are antenna-like organelles that regulate growth and development via extracellular signals. However, the molecular mechanisms underlying cilia dynamics, particularly those regulating their disassembly, are not well understood. Here, we show that leucine-rich repeat kinase 1 (LRRK1) plays a role in regulating cilia disassembly. The depletion of LRRK1 impairs primary cilia resorption following serum stimulation in cultured cells. Polo-like kinase 1 (PLK1) plays an important role in this process. During ciliary resorption, PLK1 phosphorylates LRRK1 at the primary cilia base, resulting in its activation. We identified nuclear distribution protein nudE-like 1 (NDEL1), which is known to positively regulate cilia disassembly, as a target of LRRK1 phosphorylation. Whereas LRRK1 phosphorylation of NDEL1 on Ser-155 promotes NDEL1 interaction with the intermediate chains of cytoplasmic dynein-2, it is also crucial for triggering ciliary resorption through dynein-2-driven retrograde intraflagellar transport. These findings provide evidence that a novel PLK1–LRRK1–NDEL1 pathway regulates cilia disassembly.
- Published
- 2022
6. Histidine dephosphorylation of the Gβ protein <scp>GPB</scp> ‐1 promotes axon regeneration in C. elegans
- Author
-
Yoshiki Sakai, Hiroshi Hanafusa, Naoki Hisamoto, and Kunihiro Matsumoto
- Subjects
Genetics ,Molecular Biology ,Biochemistry - Abstract
Histidine phosphorylation is an emerging noncanonical protein phosphorylation in animals, yet its physiological role remains largely unexplored. The protein histidine phosphatase (PHPT1) was recently identified for the first time in mammals. Here, we report that PHIP-1, an ortholog of PHPT1 in Caenorhabditis elegans, promotes axon regeneration by dephosphorylating GPB-1 Gβ at His-266 and inactivating GOA-1 Goα signaling, a negative regulator of axon regeneration. Overexpression of the histidine kinase NDK-1 also inhibits axon regeneration via GPB-1 His-266 phosphorylation. Thus, His-phosphorylation plays an antiregenerative role in C. elegans. Furthermore, we identify a conserved UNC-51/ULK kinase that functions in autophagy as a PHIP-1-binding protein. We demonstrate that UNC-51 phosphorylates PHIP-1 at Ser-112 and activates its catalytic activity and that this phosphorylation is required for PHIP-1-mediated axon regeneration. This study reveals a molecular link from ULK to protein histidine phosphatase, which facilitates axon regeneration by inhibiting trimeric G protein signaling.
- Published
- 2022
7. BRCA1–BARD1 Regulates Axon Regeneration in Concert with the Gqα–DAG Signaling Network
- Author
-
Hiroshi Hanafusa, Naoki Hisamoto, Strahil Iv. Pastuhov, Tatsuhiro Shimizu, Kunihiro Matsumoto, and Yoshiki Sakai
- Subjects
0301 basic medicine ,Diacylglycerol Kinase ,Ubiquitin-Protein Ligases ,BRCA1-BARD1 ,Animals, Genetically Modified ,03 medical and health sciences ,0302 clinical medicine ,BARD1 ,Chlorocebus aethiops ,medicine ,Animals ,Amino Acid Sequence ,Axon ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Mitosis ,Research Articles ,Diacylglycerol kinase ,biology ,Chemistry ,Tumor Suppressor Proteins ,General Neuroscience ,Regeneration (biology) ,axon regeneration ,biology.organism_classification ,Axons ,Nerve Regeneration ,Cell biology ,Ubiquitin ligase ,030104 developmental biology ,medicine.anatomical_structure ,COS Cells ,biology.protein ,GTP-Binding Protein alpha Subunits, Gq-G11 ,Signal transduction ,030217 neurology & neurosurgery ,Signal Transduction ,Cellular/Molecular - Abstract
The breast cancer susceptibility protein BRCA1 and its partner BRCA1-associated RING domain protein 1 (BARD1) form an E3-ubiquitin (Ub) ligase complex that acts as a tumor suppressor in mitotic cells. However, the roles of BRCA1–BARD1 in postmitotic cells, such as neurons, remain poorly defined. Here, we report that BRC-1 and BRD-1, theCaenorhabditis elegansorthologs of BRCA1 and BARD1, are required for adult-specific axon regeneration, which is positively regulated by the EGL-30 Gqα–diacylglycerol (DAG) signaling pathway. This pathway is downregulated by DAG kinase (DGK), which converts DAG to phosphatidic acid (PA). We demonstrate that inactivation of DGK-3 suppresses thebrc-1 brd-1defect in axon regeneration, suggesting that BRC-1–BRD-1 inhibits DGK-3 function. Indeed, we show that BRC-1–BRD-1 poly-ubiquitylates DGK-3 in a manner dependent on its E3 ligase activity, causing DGK-3 degradation. Furthermore, we find that axon injury causes the translocation of BRC-1 from the nucleus to the cytoplasm, where DGK-3 is localized. These results suggest that the BRC-1–BRD-1 complex regulates axon regeneration in concert with the Gqα–DAG signaling network. Thus, this study describes a new role for breast cancer proteins in fully differentiated neurons and the molecular mechanism underlying the regulation of axon regeneration in response to nerve injury.SIGNIFICANCE STATEMENTBRCA1–BRCA1-associated RING domain protein 1 (BARD1) is an E3-ubiquitin (Ub) ligase complex acting as a tumor suppressor in mitotic cells. The roles of BRCA1–BARD1 in postmitotic cells, such as neurons, remain poorly defined. We show here thatCaenorhabditis elegansBRC-1/BRCA1 and BRD-1/BARD1 are required for adult-specific axon regeneration, a process that requires high diacylglycerol (DAG) levels in injured neurons. The DAG kinase (DGK)-3 inhibits axon regeneration by reducing DAG levels. We find that BRC-1–BRD-1 poly-ubiquitylates and degrades DGK-3, thereby keeping DAG levels elevated and promoting axon regeneration. Furthermore, we demonstrate that axon injury causes the translocation of BRC-1 from the nucleus to the cytoplasm, where DGK-3 is localized. Thus, this study describes a new role for BRCA1–BARD1 in fully-differentiated neurons.
- Published
- 2021
8. Caenorhabditis elegansF-Box Protein Promotes Axon Regeneration by Inducing Degradation of the Mad Transcription Factor
- Author
-
Hiroshi Hanafusa, Yoshiki Sakai, Naoki Hisamoto, Strahil Iv. Pastuhov, Yasuko Todoroki, Tatsuhiro Shimizu, and Kunihiro Matsumoto
- Subjects
0301 basic medicine ,F-box protein ,Receptor tyrosine kinase ,03 medical and health sciences ,0302 clinical medicine ,Mad/Max ,ubiquitin ,Gene expression ,medicine ,Axon ,Transcription factor ,Research Articles ,Caenorhabditis elegans ,biology ,Chemistry ,General Neuroscience ,Regeneration (biology) ,axon regeneration ,biology.organism_classification ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,nervous system ,Proteasome ,C. elegans ,biology.protein ,030217 neurology & neurosurgery ,Cellular/Molecular - Abstract
In Caenorhabditis elegans, axon regeneration is activated by a signaling cascade through the receptor tyrosine kinase (RTK) SVH-2. Axonal injury induces svh-2 gene expression by degradation of the Mad-like transcription factor MDL-1. In this study, we identify the svh-24/sdz-33 gene encoding a protein containing F-box and F-box-associated domains as a regulator of axon regeneration in motor neurons. We find that sdz-33 is required for axon injury-induced svh-2 expression., In Caenorhabditis elegans, axon regeneration is activated by a signaling cascade through the receptor tyrosine kinase (RTK) SVH-2. Axonal injury induces svh-2 gene expression by degradation of the Mad-like transcription factor MDL-1. In this study, we identify the svh-24/sdz-33 gene encoding a protein containing F-box and F-box-associated domains as a regulator of axon regeneration in motor neurons. We find that sdz-33 is required for axon injury-induced svh-2 expression. SDZ-33 targets MDL-1 for poly-ubiquitylation and degradation. Furthermore, we demonstrate that SDZ-33 promotes axotomy-induced nuclear degradation of MDL-1, resulting in the activation of svh-2 expression in animals. These results suggest that the F-box protein is required for RTK signaling in the control of axon regeneration. SIGNIFICANCE STATEMENT In Caenorhabditis elegans, axon regeneration is positively regulated by the growth factor SVH-1 and its receptor tyrosine kinase SVH-2. Expression of the svh-2 gene is induced by axonal injury via the Ets-like transcription factor ETS-4, whose transcriptional activity is inhibited by the Mad-like transcription factor MDL-1. Axon injury leads to the degradation of MDL-1, and this is linked to the activation of ETS-4 transcriptional activity. In this study, we identify the sdz-33 gene encoding a protein containing an F-box domain as a regulator of axon regeneration. We demonstrate that MDL-1 is poly-ubiquitylated and degraded through the SDZ-33-mediated 26S proteasome pathway. These results reveal that an F-box protein promotes axon regeneration by degrading the Mad transcription factor.
- Published
- 2021
9. The Integrin Signaling Network Promotes Axon Regeneration via the Src–Ephexin–RhoA GTPase Signaling Axis
- Author
-
Mayuka Tsunekawa, Yoshiki Sakai, Naoki Hisamoto, Hiroshi Hanafusa, Kohei Ohta, Kunihiro Matsumoto, Strahil Iv. Pastuhov, and Tatsuhiro Shimizu
- Subjects
0301 basic medicine ,Integrins ,RHOA ,integrin ,Integrin ,GTPase ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Guanine Nucleotide Exchange Factors ,Axon ,Caenorhabditis elegans ,Research Articles ,biology ,Chemistry ,inside-out ,General Neuroscience ,axon regeneration ,RhoA ,Axons ,Cell biology ,Nerve Regeneration ,030104 developmental biology ,medicine.anatomical_structure ,src-Family Kinases ,biology.protein ,C. elegans ,Phosphorylation ,Guanine nucleotide exchange factor ,Signal transduction ,rhoA GTP-Binding Protein ,030217 neurology & neurosurgery ,Proto-oncogene tyrosine-protein kinase Src ,Cellular/Molecular ,Src ,Signal Transduction - Abstract
Axon regeneration is an evolutionarily conserved process essential for restoring the function of damaged neurons. InCaenorhabditis eleganshermaphrodites, initiation of axon regeneration is regulated by the RhoA GTPase–ROCK (Rho-associated coiled-coil kinase)–regulatory nonmuscle myosin light-chain phosphorylation signaling pathway. However, the upstream mechanism that activates the RhoA pathway remains unknown. Here, we show that axon injury activates TLN-1/talin via the cAMP–Epac (exchange protein directly activated by cAMP)–Rap GTPase cascade and that TLN-1 induces multiple downstream events, one of which is integrin inside-out activation, leading to the activation of the RhoA–ROCK signaling pathway. We found that the nonreceptor tyrosine kinase Src, a key mediator of integrin signaling, activates the Rho guanine nucleotide exchange factor EPHX-1/ephexin by phosphorylating the Tyr-568 residue in the autoinhibitory domain. Our results suggest that theC. elegansintegrin signaling network regulates axon regeneration via the Src–RhoGEF–RhoA axis.SIGNIFICANCE STATEMENTThe ability of axons to regenerate after injury is governed by cell-intrinsic regeneration pathways. We have previously demonstrated that theCaenorhabditis elegansRhoA GTPase–ROCK (Rho-associated coiled-coil kinase) pathway promotes axon regeneration by inducing MLC-4 phosphorylation. In this study, we found that axon injury activates TLN-1/talin through the cAMP–Epac (exchange protein directly activated by cAMP)–Rap GTPase cascade, leading to integrin inside-out activation, which promotes axonal regeneration by activating the RhoA signaling pathway. In this pathway, SRC-1/Src acts downstream of integrin activation and subsequently activates EPHX-1/ephexin RhoGEF by phosphorylating the Tyr-568 residue in the autoinhibitory domain. Our results suggest that theC. elegansintegrin signaling network regulates axon regeneration via the Src–RhoGEF–RhoA axis.
- Published
- 2021
10. A fast search method of similar strings from dictionaries.
- Author
-
Masao Fuketa, El-Sayed Atlam, Nobuo Fujisawa, Hiroshi Hanafusa, Kazuhiro Morita, and Jun-ichi Aoe
- Published
- 2011
- Full Text
- View/download PDF
11. A method of extracting malicious expressions in bulletin board systems by using context analysis.
- Author
-
Hiroshi Hanafusa, Kazuhiro Morita, Masao Fuketa, and Jun-ichi Aoe
- Published
- 2011
- Full Text
- View/download PDF
12. CDK14 Promotes Axon Regeneration by Regulating the Noncanonical Wnt Signaling Pathway in a Kinase-Independent Manner
- Author
-
Tatsuhiro Shimizu, Hiroshi Hanafusa, Naoki Hisamoto, Yoshiki Sakai, Kunihiro Matsumoto, Kohei Ohta, and Chun Li
- Subjects
Frizzled ,GTPase ,macromolecular substances ,Animals, Genetically Modified ,Chlorocebus aethiops ,medicine ,Animals ,Axon ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,noncanonical Wnt signaling pathway ,Wnt Signaling Pathway ,Research Articles ,Chemistry ,General Neuroscience ,Regeneration (biology) ,Wnt signaling pathway ,axon regeneration ,Axons ,Cyclin-Dependent Kinases ,Cell biology ,Nerve Regeneration ,medicine.anatomical_structure ,cdk14 ,COS Cells ,C. elegans ,Phosphorylation ,Guanine nucleotide exchange factor ,Signal transduction ,Cellular/Molecular - Abstract
The postinjury regenerative capacity of neurons is known to be mediated by a complex interaction of intrinsic regenerative pathways and external cues. In Caenorhabditis elegans, the initiation of axon regeneration is regulated by the nonmuscle myosin light chain-4 (MLC-4) phosphorylation signaling pathway. In this study, we have identified svh-16/cdk-14, a mammalian CDK14 homolog, as a positive regulator of axon regeneration in motor neurons. We then isolated the CDK-14-binding protein MIG-5/Disheveled (Dsh) and found that EGL-20/Wnt and the MIG-1/Frizzled receptor (Fz) are required for efficient axon regeneration. Further, we demonstrate that CDK-14 activates EPHX-1, the C. elegans homolog of the mammalian ephexin Rho-type GTPase guanine nucleotide exchange factor (GEF), in a kinase-independent manner. EPHX-1 functions as a GEF for the CDC-42 GTPase, inhibiting myosin phosphatase, which maintains MLC-4 phosphorylation. These results suggest that CDK14 activates the RhoGEF–CDC42–MLC phosphorylation axis in a noncanonical Wnt signaling pathway that promotes axon regeneration. SIGNIFICANCE STATEMENT Noncanonical Wnt signaling is mediated by Frizzled receptor (Fz), Disheveled (Dsh), Rho-type GTPase, and nonmuscle myosin light chain (MLC) phosphorylation. This study identified svh-16/cdk-14, which encodes a mammalian CDK14 homolog, as a regulator of axon regeneration in Caenorhabditis elegans motor neurons. We show that CDK-14 binds to MIG-5/Dsh, and that EGL-20/Wnt, MIG-1/Fz, and EPHX-1/RhoGEF are required for axon regeneration. The phosphorylation-mimetic MLC-4 suppressed axon regeneration defects in mig-1, cdk-14, and ephx-1 mutants. CDK-14 mediates kinase-independent activation of EPHX-1, which functions as a guanine nucleotide exchange factor for CDC-42 GTPase. Activated CDC-42 inactivates myosin phosphatase and thereby maintains MLC phosphorylation. Thus, the noncanonical Wnt signaling pathway controls axon regeneration via the CDK-14–EPHX-1–CDC-42–MLC phosphorylation axis.
- Published
- 2021
13. TDP2 negatively regulates axon regeneration by inducing SUMOylation of an Ets transcription factor
- Author
-
Kunihiro Matsumoto, Hiroshi Hanafusa, Chun Li, Strahil Iv. Pastuhov, Tatsuhiro Shimizu, Naoki Hisamoto, and Yoshiki Sakai
- Subjects
MAPK/ERK pathway ,Transcription, Genetic ,SUMO protein ,Biochemistry ,Models, Biological ,Receptor tyrosine kinase ,Article ,03 medical and health sciences ,0302 clinical medicine ,TDP2/EAPII ,Genetics ,medicine ,Animals ,Axon ,Phosphorylation ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Molecular Biology ,Transcription factor ,030304 developmental biology ,Motor Neurons ,0303 health sciences ,biology ,Proto-Oncogene Proteins c-ets ,Chemistry ,Phosphoric Diester Hydrolases ,Regeneration (biology) ,ETS transcription factor family ,axon regeneration ,Post-translational Modifications, Proteolysis & Proteomics ,Sumoylation ,Articles ,C. elegans ,Axons ,Cell biology ,Nerve Regeneration ,DNA-Binding Proteins ,medicine.anatomical_structure ,Gene Expression Regulation ,Mitogen-activated protein kinase ,biology.protein ,030217 neurology & neurosurgery ,Ets ,Neuroscience ,Protein Binding ,Transcription Factors - Abstract
In Caenorhabditis elegans, the JNK MAP kinase (MAPK) pathway is important for axon regeneration. The JNK pathway is activated by a signaling cascade consisting of the growth factor SVH‐1 and its receptor tyrosine kinase SVH‐2. Expression of the svh‐2 gene is induced by axonal injury in a process involving the transcription factors ETS‐4 and CEBP‐1. Here, we find that svh‐14/mxl‐1, a gene encoding a Max‐like transcription factor, is required for activation of svh‐2 expression in response to axonal injury. We show that MXL‐1 binds to and inhibits the function of TDPT‐1, a C. elegans homolog of mammalian tyrosyl‐DNA phosphodiesterase 2 [TDP2; also called Ets1‐associated protein II (EAPII)]. Deletion of tdpt‐1 suppresses the mxl‐1 defect, but not the ets‐4 defect, in axon regeneration. TDPT‐1 induces SUMOylation of ETS‐4, which inhibits ETS‐4 transcriptional activity, and MXL‐1 counteracts this effect. Thus, TDPT‐1 interacts with two different transcription factors in axon regeneration.
- Published
- 2019
14. C. elegans Tensin Promotes Axon Regeneration by Linking the Met-like SVH-2 and Integrin Signaling Pathways
- Author
-
Kazuma Asai, Yoshiki Sakai, Tatsuhiro Shimizu, Hiroshi Hanafusa, Naoki Hisamoto, Strahil Iv. Pastuhov, and Kunihiro Matsumoto
- Subjects
0301 basic medicine ,biology ,Chemistry ,General Neuroscience ,Regeneration (biology) ,SH2 domain ,Receptor tyrosine kinase ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,nervous system ,Mitogen-activated protein kinase ,medicine ,biology.protein ,Tensin ,Axon ,Signal transduction ,Phosphotyrosine-binding domain ,030217 neurology & neurosurgery ,Research Articles - Abstract
Axon regeneration is a conserved mechanism induced by axon injury that initiates a neuronal response leading to regrowth of the axon. In Caenorhabditis elegans, the initiation of axon regeneration is regulated by the JNK MAP kinase (MAPK) pathway. We have previously identified a number of genes affecting the JNK pathway using an RNAi-based screen. Analysis of these genes, called the svh genes, has shed new light on the regulation of axon regeneration, revealing the involvement of a signaling cascade consisting of a growth factor SVH-1 and its receptor, the tyrosine kinase SVH-2. Here, we characterize the svh-6/tns-1 gene, which is a homolog of mammalian tensin, and show that it is a positive regulator of axon regeneration in motor neurons. We demonstrate that TNS-1 interacts with tyrosine-autophosphorylated SVH-2 and the integrin β subunit PAT-3 via its SH2 and PTB domains, respectively, to promote axon regeneration. These results suggest that TNS-1 acts as an adaptor to link the SVH-2 and integrin signaling pathways. SIGNIFICANCE STATEMENT The Caenorhabditis elegans JNK MAPK pathway regulates the initiation of axon regeneration. Previously, we showed that a signaling cascade consisting of the HGF-like growth factor SVH-1 and its Met-like receptor tyrosine kinase SVH-2 promotes axon regeneration through activation of the JNK pathway. In this study, we show that the C. elegans tensin, TNS-1, is required for efficient regeneration after axon injury. Phosphorylation of SVH-2 on tyrosine mediates its interaction with the SH2 domain of TNS-1 to positively regulate axon regeneration. Furthermore, TNS-1 interacts via its PTB domain with the integrin β subunit PAT-3. These results suggest that TNS-1 plays a critical role in the regulation of axon regeneration by linking the SVH-2 and integrin signaling pathways.
- Published
- 2019
15. LRRK1 phosphorylation of Rab7 at S72 links trafficking of EGFR-containing endosomes to its effector RILP
- Author
-
Hiroshi Hanafusa, Kozo Kaibuchi, Naoki Hisamoto, Kyoko Shirakabe, Kunihiro Matsumoto, Haruka Ikeda, Tomoki Nishioka, and Takuya Yagi
- Subjects
Endosome ,Endocytic cycle ,macromolecular substances ,Endosomes ,Protein Serine-Threonine Kinases ,Cell Line ,Motor protein ,03 medical and health sciences ,0302 clinical medicine ,Chlorocebus aethiops ,Animals ,Humans ,Epidermal growth factor receptor ,Phosphorylation ,030304 developmental biology ,Adaptor Proteins, Signal Transducing ,0303 health sciences ,biology ,Effector ,Dyneins ,rab7 GTP-Binding Proteins ,Cell Biology ,Dynactin Complex ,Cell biology ,ErbB Receptors ,Actin Cytoskeleton ,Protein Transport ,HEK293 Cells ,RAB7A ,rab GTP-Binding Proteins ,COS Cells ,biology.protein ,030217 neurology & neurosurgery ,Intracellular ,HeLa Cells - Abstract
Ligand-induced activation of epidermal growth factor receptor (EGFR) initiates trafficking events that re-localize the receptor from the cell surface to intracellular endocytic compartments. EGFR-containing endosomes are transported to lysosomes for degradation by the dynein-dynactin motor protein complex. However, this cargo-dependent endosomal trafficking mechanism remains largely uncharacterized. Here, we show that GTP-bound Rab7 is phosphorylated on S72 by leucine-rich repeat kinase 1 (LRRK1) at the endosomal membrane. This phosphorylation promotes the interaction of Rab7 (herein referring to Rab7a) with its effector RILP, resulting in recruitment of the dynein-dynactin complex to Rab7-positive vesicles. This, in turn, facilitates the dynein-driven transport of EGFR-containing endosomes toward the perinuclear region. These findings reveal a mechanism regulating the cargo-specific trafficking of endosomes.
- Published
- 2018
16. CDK14 Promotes Axon Regeneration by Regulating the Noncanonical Wnt Signaling Pathway in a Kinase- Independent Manner.
- Author
-
Naoki Hisamoto, Yoshiki Sakai, Kohei Ohta, Tatsuhiro Shimizu, Chun Li, Hiroshi Hanafusa, and Kunihiro Matsumoto
- Subjects
CELLULAR signal transduction ,WNT signal transduction ,GUANINE nucleotide exchange factors ,NERVOUS system regeneration ,MYOSIN ,AXONS ,CHONDROITIN sulfate proteoglycan - Abstract
The postinjury regenerative capacity of neurons is known to be mediated by a complex interaction of intrinsic regenerative pathways and external cues. In Caenorhabditis elegans, the initiation of axon regeneration is regulated by the nonmuscle myosin light chain-4 (MLC-4) phosphorylation signaling pathway. In this study, we have identified svh-16/cdk-14, a mammalian CDK14 homolog, as a positive regulator of axon regeneration in motor neurons. We then isolated the CDK-14-binding protein MIG-5/Disheveled (Dsh) and found that EGL-20/Wnt and the MIG-1/Frizzled receptor (Fz) are required for efficient axon regeneration. Further, we demonstrate that CDK-14 activates EPHX-1, the C. elegans homolog of the mammalian ephexin Rho-type GTPase guanine nucleotide exchange factor (GEF), in a kinase-independent manner. EPHX-1 functions as a GEF for the CDC-42 GTPase, inhibiting myosin phosphatase, which maintains MLC-4 phosphorylation. These results suggest that CDK14 activates the RhoGEF–CDC42–MLC phosphorylation axis in a noncanonical Wnt signaling pathway that promotes axon regeneration. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
17. Chaperone complex BAG2-HSC70 regulates localization ofCaenorhabditis elegansleucine-rich repeat kinase LRK-1 to the Golgi
- Author
-
Hiroshi Hanafusa, Shun-ichiro Iemura, Hiroshi Shibuya, Strahil Iv. Pastuhov, Tohru Natsume, Okinobu Fukushima, Ayuna Hattori, Takashi Fukuzono, Naoki Hisamoto, Chun Li, and Kunihiro Matsumoto
- Subjects
0301 basic medicine ,Mutant ,Golgi Apparatus ,Protein Serine-Threonine Kinases ,Leucine-rich repeat ,Synaptic vesicle ,03 medical and health sciences ,symbols.namesake ,0302 clinical medicine ,Genetics ,Animals ,HSP70 Heat-Shock Proteins ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,biology ,Cell Biology ,Golgi apparatus ,biology.organism_classification ,LRRK2 ,Phenotype ,Cell biology ,030104 developmental biology ,symbols ,Chaperone complex ,Synaptic Vesicles ,Carrier Proteins ,030217 neurology & neurosurgery ,Molecular Chaperones - Abstract
Mutations in LRRK2 are linked to autosomal dominant forms of Parkinson's disease. We identified two human proteins that bind to LRRK2: BAG2 and HSC70, which are known to form a chaperone complex. We characterized the role of their Caenorhabditis elegans homologues, UNC-23 and HSP-1, in the regulation of LRK-1, the sole homologue of human LRRK2. In C. elegans, LRK-1 determines the polarized sorting of synaptic vesicle (SV) proteins to the axons by excluding SV proteins from the dendrite-specific transport machinery in the Golgi. In unc-23 mutants, SV proteins are localized to both presynaptic and dendritic endings in neurons, a phenotype also observed in lrk-1 deletion mutants. Furthermore, we isolated mutations in the hsp-1 gene that can suppress the unc-23, but not the lrk-1 defect. We show that UNC-23 determines LRK-1 localization to the Golgi apparatus in cooperation with HSP-1. These results describe a chaperone-dependent mechanism through which LRK-1 localization is regulated.
- Published
- 2016
18. The Integrin Signaling Network Promotes Axon Regeneration via the Src-Ephexin-RhoA GTPase Signaling Axis.
- Author
-
Yoshiki Sakai, Mayuka Tsunekawa, Kohei Ohta, Tatsuhiro Shimizu, Pastuhov, Strahil Iv., Hiroshi Hanafusa, Naoki Hisamoto, and Kunihiro Matsumoto
- Subjects
RHO-associated kinases ,SMOOTH muscle ,GUANINE nucleotide exchange factors ,NERVOUS system regeneration ,GUANOSINE triphosphatase ,INTEGRINS - Abstract
Axon regeneration is an evolutionarily conserved process essential for restoring the function of damaged neurons. In Caenorhabditis elegans hermaphrodites, initiation of axon regeneration is regulated by the RhoA GTPase-ROCK (Rho-associated coiled-coil kinase)-regulatory nonmuscle myosin light-chain phosphorylation signaling pathway. However, the upstream mechanism that activates the RhoA pathway remains unknown. Here, we show that axon injury activates TLN-1/talin via the cAMP-Epac (exchange protein directly activated by cAMP)-Rap GTPase cascade and that TLN-1 induces multiple downstream events, one of which is integrin inside-out activation, leading to the activation of the RhoA-ROCK signaling pathway. We found that the nonreceptor tyrosine kinase Src, a key mediator of integrin signaling, activates the Rho guanine nucleotide exchange factor EPHX-1/ephexin by phosphorylating the Tyr-568 residue in the autoinhibitory domain. Our results suggest that the C. elegans integrin signaling network regulates axon regeneration via the Src-RhoGEF-RhoA axis. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
19. [Regulation of cellular function by ROCO family kinase LRRK1 in a manner dependent on its substrates]
- Author
-
Hiroshi, Hanafusa and Kunihiro, Matsumoto
- Subjects
ErbB Receptors ,Protein Transport ,Animals ,M Phase Cell Cycle Checkpoints ,Spindle Apparatus ,Phosphorylation ,Protein Serine-Threonine Kinases ,Substrate Specificity - Published
- 2018
20. The C. elegans BRCA2-ALP/Enigma Complex Regulates Axon Regeneration via a Rho GTPase-ROCK-MLC Phosphorylation Pathway
- Author
-
Strahil Iv. Pastuhov, Naoki Hisamoto, Hiroshi Hanafusa, Kunihiro Matsumoto, and Tatsuhiro Shimizu
- Subjects
0301 basic medicine ,rho GTP-Binding Proteins ,Myosin Light Chains ,Neuronal Outgrowth ,GTPase ,macromolecular substances ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Myosin ,medicine ,Animals ,Axon ,Phosphorylation ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,lcsh:QH301-705.5 ,Adaptor Proteins, Signal Transducing ,Motor Neurons ,rho-Associated Kinases ,Neuronal Plasticity ,biology ,Chemistry ,Kinase ,Regeneration (biology) ,Axotomy ,LIM Domain Proteins ,biology.organism_classification ,musculoskeletal system ,Cell biology ,Nerve Regeneration ,DNA-Binding Proteins ,030104 developmental biology ,medicine.anatomical_structure ,lcsh:Biology (General) ,Gene Expression Regulation ,Signal transduction ,Protein Binding ,Signal Transduction - Abstract
Summary: The ability of specific neurons to regenerate their axons after injury is governed by cell-intrinsic regeneration pathways. However, the mechanisms regulating axon regeneration are not well understood. Here, we identify the brc-2 gene encoding a homolog of the mammalian BRCA2 tumor suppressor as a regulator of axon regeneration in Caenorhabditis elegans motor neurons. We show that the RHO-1/Rho GTPase-LET-502/ROCK (Rho-associated coiled-coil kinase)-regulatory non-muscle myosin light-chain (MLC-4/MLC) phosphorylation signaling pathway regulates axon regeneration. BRC-2 functions between RHO-1 and LET-502, suggesting that BRC-2 is required for the activation of LET-502 by RHO-1-GTP. We also find that one component that interacts with BRC-2, the ALP (α-actinin-associated LIM protein)/Enigma protein ALP-1, is required for regeneration and acts between LET-502 and MLC-4 phosphorylation. Furthermore, we demonstrate that ALP-1 associates with LET-502 and MLC-4. Thus, ALP-1 serves as a platform to activate MLC-4 phosphorylation mediated by the RHO-1–LET-502 signaling pathway. : Shimizu et al. demonstrate that a BRCA2 homolog BRC-2 and its binding partner, the ALP/Enigma protein ALP-1, regulate axon regeneration via the Rho-ROCK-MLC signaling pathway in C. elegans. ALP-1 serves as a platform to activate MLC phosphorylation, which promotes growth cone formation of severed axons. Keywords: axon regeneration, BRCA2, Rho kinase, C. elegans
- Published
- 2017
21. PLK1-dependent activation of LRRK1 regulates spindle orientation by phosphorylating CDK5RAP2
- Author
-
Satoshi Usami, Kunihiro Matsumoto, Hiroshi Hanafusa, Motohiro Tezuka, Fumiko Toyoshima, Shin Kedashiro, and Motoki Funatsu
- Subjects
inorganic chemicals ,Time Factors ,Amino Acid Motifs ,Mitosis ,Cell Cycle Proteins ,Nerve Tissue Proteins ,Spindle Apparatus ,macromolecular substances ,Protein Serine-Threonine Kinases ,Editorials: Cell Cycle Features ,Transfection ,Microtubules ,environment and public health ,PLK1 ,Spindle pole body ,Enzyme activator ,Tubulin ,Proto-Oncogene Proteins ,CDC2 Protein Kinase ,Chlorocebus aethiops ,Serine ,Animals ,Humans ,Phosphorylation ,Microtubule nucleation ,Centrosome ,Binding Sites ,COS cells ,Chemistry ,Intracellular Signaling Peptides and Proteins ,Cell Biology ,Cyclin-Dependent Kinases ,Cell biology ,Enzyme Activation ,enzymes and coenzymes (carbohydrates) ,HEK293 Cells ,COS Cells ,Mutation ,bacteria ,RNA Interference ,HeLa Cells ,Protein Binding ,Signal Transduction - Abstract
Correct formation of the cell division axis requires the initial precise orientation of the mitotic spindle. Proper spindle orientation depends on centrosome maturation, and Polo-like kinase 1 (PLK1) is known to play a crucial role in this process. However, the molecular mechanisms that function downstream of PLK1 are not well understood. Here we show that LRRK1 is a PLK1 substrate that is phosphorylated on Ser 1790. PLK1 phosphorylation is required for CDK1-mediated activation of LRRK1 at the centrosomes, and this in turn regulates mitotic spindle orientation by nucleating the growth of astral microtubules from the centrosomes. Interestingly, LRRK1 in turn phosphorylates CDK5RAP2(Cep215), a human homologue of Drosophila Centrosomin (Cnn), in its γ-tubulin-binding motif, thus promoting the interaction of CDK5RAP2 with γ-tubulin. LRRK1 phosphorylation of CDK5RAP2 Ser 140 is necessary for CDK5RAP2-dependent microtubule nucleation. Thus, our findings provide evidence that LRRK1 regulates mitotic spindle orientation downstream of PLK1 through CDK5RAP2-dependent centrosome maturation.
- Published
- 2015
22. BRCA1-BARD1 Regulates Axon Regeneration in Concert with the Gqα-DAG Signaling Network.
- Author
-
Yoshiki Sakai, Hiroshi Hanafusa, Tatsuhiro Shimizu, Pastuhov, Strahil I., Naoki Hisamoto, and Kunihiro Matsumoto
- Subjects
- *
NERVOUS system regeneration , *AXONS , *SUPPRESSOR cells , *CAENORHABDITIS elegans ,CANCER susceptibility - Abstract
The breast cancer susceptibility protein BRCA1 and its partner BRCA1-associated RING domain protein 1 (BARD1) form an E3-ubiquitin (Ub) ligase complex that acts as a tumor suppressor in mitotic cells. However, the roles of BRCA1-BARD1 in postmitotic cells, such as neurons, remain poorly defined. Here, we report that BRC-1 and BRD-1, the Caenorhabditis elegans orthologs of BRCA1 and BARD1, are required for adult-specific axon regeneration, which is positively regulated by the EGL-30 Gqα-diacylglycerol (DAG) signaling pathway. This pathway is downregulated by DAG kinase (DGK), which converts DAG to phosphatidic acid (PA). We demonstrate that inactivation of DGK-3 suppresses the brc-1 brd-1 defect in axon regeneration, suggesting that BRC-1-BRD-1 inhibits DGK-3 function. Indeed, we show that BRC-1-BRD-1 poly-ubiquitylates DGK-3 in a manner dependent on its E3 ligase activity, causing DGK-3 degradation. Furthermore, we find that axon injury causes the translocation of BRC-1 from the nucleus to the cytoplasm, where DGK-3 is localized. These results suggest that the BRC-1-BRD-1 complex regulates axon regeneration in concert with the Gqα-DAG signaling network. Thus, this study describes a new role for breast cancer proteins in fully differentiated neurons and the molecular mechanism underlying the regulation of axon regeneration in response to nerve injury.SIGNIFICANCE STATEMENT BRCA1-BRCA1-associated RING domain protein 1 (BARD1) is an E3-ubiquitin (Ub) ligase complex acting as a tumor suppressor in mitotic cells. The roles of BRCA1-BARD1 in postmitotic cells, such as neurons, remain poorly defined. We show here that Caenorhabditis elegans BRC-1/BRCA1 and BRD-1/BARD1 are required for adult-specific axon regeneration, a process that requires high diacylglycerol (DAG) levels in injured neurons. The DAG kinase (DGK)-3 inhibits axon regeneration by reducing DAG levels. We find that BRC-1-BRD-1 poly-ubiquitylates and degrades DGK-3, thereby keeping DAG levels elevated and promoting axon regeneration. Furthermore, we demonstrate that axon injury causes the translocation of BRC-1 from the nucleus to the cytoplasm, where DGK-3 is localized. Thus, this study describes a new role for BRCA1-BARD1 in fully-differentiated neurons. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
23. Caenorhabditis elegans F-Box Protein Promotes Axon Regeneration by Inducing Degradation of the Mad Transcription Factor.
- Author
-
Tatsuhiro Shimizu, Pastuhov, Strahil I., Hiroshi Hanafusa, Yoshiki Sakai, Yasuko Todoroki, Naoki Hisamoto, and Kunihiro Matsumoto
- Subjects
CAENORHABDITIS elegans ,TRANSCRIPTION factors ,NERVOUS system regeneration ,AXONS ,MOTOR neurons - Abstract
In Caenorhabditis elegans, axon regeneration is activated by a signaling cascade through the receptor tyrosine kinase (RTK) SVH-2. Axonal injury induces svh-2 gene expression by degradation of the Mad-like transcription factor MDL-1. In this study, we identify the svh-24/sdz-33 gene encoding a protein containing F-box and F-box-associated domains as a regulator of axon regeneration in motor neurons. We find that sdz-33 is required for axon injury-induced svh-2 expression. SDZ-33 targets MDL-1 for poly-ubiquitylation and degradation. Furthermore, we demonstrate that SDZ-33 promotes axotomy-induced nuclear degradation of MDL-1, resulting in the activation of svh-2 expression in animals. These results suggest that the F-box protein is required for RTK signaling in the control of axon regeneration. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
24. Phosphatidylserine exposure mediated by ABC transporter activates the integrin signaling pathway promoting axon regeneration
- Author
-
Naoki Hisamoto, Tatsuhiro Shimizu, Hiroshi Hanafusa, Strahil Iv. Pastuhov, Anna Tsuge, and Kunihiro Matsumoto
- Subjects
0301 basic medicine ,Integrins ,General Physics and Astronomy ,ATP-binding cassette transporter ,Apoptosis ,Animals, Genetically Modified ,chemistry.chemical_compound ,0302 clinical medicine ,Axon ,lcsh:Science ,Caspase ,Multidisciplinary ,biology ,Phosphatidylserine ,Lipids ,Cell biology ,rac GTP-Binding Proteins ,medicine.anatomical_structure ,Caspases ,Signal transduction ,Plasmids ,Signal Transduction ,endocrine system ,Science ,Integrin ,Phosphatidylserines ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,Phagocytosis ,medicine ,Animals ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Integrin Signaling Pathway ,Regeneration (biology) ,fungi ,nutritional and metabolic diseases ,General Chemistry ,Axons ,Nerve Regeneration ,Cytoskeletal Proteins ,030104 developmental biology ,chemistry ,nervous system ,Mutation ,biology.protein ,lcsh:Q ,ATP-Binding Cassette Transporters ,030217 neurology & neurosurgery - Abstract
Following axon injury, a cascade of signaling events is triggered to initiate axon regeneration. However, the mechanisms regulating axon regeneration are not well understood at present. In Caenorhabditis elegans, axon regeneration utilizes many of the components involved in phagocytosis, including integrin and Rac GTPase. Here, we identify the transthyretin (TTR)-like protein TTR-11 as a component functioning in axon regeneration upstream of integrin. We show that TTR-11 binds to both the extracellular domain of integrin-α and phosphatidylserine (PS). Axon injury induces the accumulation of PS around the injured axons in a manner dependent on TTR-11, the ABC transporter CED-7, and the caspase CED-3. Furthermore, we demonstrate that CED-3 activates CED-7 during axon regeneration. Thus, TTR-11 functions to link the PS injury signal to activation of the integrin pathway, which then initiates axon regeneration., Apoptotic cells display surface signals such as phosphatidlyserines that are recognized by phagocytes via engulfment signal receptors. Here, the authors show how one such receptor, transthyretin-like protein 11, plays a role in initiating axon regeneration in the peripheral nervous system.
- Published
- 2017
25. TAK1 determines susceptibility to endoplasmic reticulum stress and hypothalamic leptin resistance
- Author
-
Hiroshi Hanafusa, Sho Morioka, H. Troy Ghashghaei, Nagendran Muthusamy, Kazuhito Sai, Kunihiro Matsumoto, Jun Ninomiya-Tsuji, and Giichi Takaesu
- Subjects
0301 basic medicine ,medicine.medical_specialty ,MAP kinase kinase kinase ,Endoplasmic reticulum ,Cellular homeostasis ,Cell Biology ,Biology ,MAP3K7 ,03 medical and health sciences ,030104 developmental biology ,Endocrinology ,Downregulation and upregulation ,Internal medicine ,medicine ,Unfolded protein response ,ASK1 ,Protein kinase A - Abstract
Sustained endoplasmic reticulum (ER) stress disrupts normal cellular homeostasis and leads to the development of many types of human diseases, including metabolic disorders. TAK1 (also known as MAP3K7) is a member of the mitogen-activated protein kinase kinase kinase (MAP3K) family and is activated by a diverse set of inflammatory stimuli. Here, we demonstrate that TAK1 regulates ER stress and metabolic signaling through modulation of lipid biogenesis. We found that deletion of Tak1 increased ER volume and facilitated ER-stress tolerance in cultured cells, which was mediated by upregulation of sterol-regulatory-element-binding protein (SREBP)-dependent lipogenesis. In the in vivo setting, central nervous system (CNS)-specific Tak1 deletion upregulated SREBP-target lipogenic genes and blocked ER stress in the hypothalamus. Furthermore, CNS-specific Tak1 deletion prevented ER-stress-induced hypothalamic leptin resistance and hyperphagic obesity under a high-fat diet (HFD). Thus, TAK1 is a crucial regulator of ER stress in vivo, which could be a target for alleviation of ER stress and its associated disease conditions.
- Published
- 2016
26. A method of extracting malicious expressions in bulletin board systems by using context analysis
- Author
-
Jun-ichi Aoe, Kazuhiro Morita, Hiroshi Hanafusa, and Masao Fuketa
- Subjects
Scheme (programming language) ,Sequence ,Information retrieval ,Computer science ,business.industry ,Word error rate ,Library and Information Sciences ,Management Science and Operations Research ,Computer security ,computer.software_genre ,Computer Science Applications ,Context analysis ,Order (business) ,Media Technology ,The Internet ,Bulletin board system ,business ,computer ,Information Systems ,computer.programming_language ,Test data - Abstract
Bulletin board systems are well-known basic services on the Internet for information frequent exchange. The convenience of bulletin boards enables us to communicate with other persons and to read the communication contents at any time. However, malicious postings about crimes are serious problems for serving companies and users. The extracting scheme of the traditional methods depends on words or a sequence of words without considering contexts of articles and, therefore, it takes a lot of human efforts to alert malicious articles. In order to reduce the human efforts, this paper presents a new filtering algorithm that can recover the error rate of false positive for non-malicious articles by using context analysis. The presented scheme builds detecting knowledge by introducing multi-attribute rules. By the experimental results for 11,019 test data, it turns out that sensitivity and specificity of the presented method become 38.7 and 24.1 (%) points higher than traditional method, respectively.
- Published
- 2011
27. Expression of Siamois and Twin in the blastula Chordin/Noggin signaling center is required for brain formation in Xenopus laevis embryos
- Author
-
Hideyuki Ishibashi, Hiroshi Hanafusa, Hiroki Kuroda, E. M. De Robertis, Noriko Matsumura, and Kunihiro Matsumoto
- Subjects
Embryology ,animal structures ,Morpholino ,Xenopus ,Biology ,Xenopus Proteins ,Article ,Xenopus laevis ,Animals ,Noggin ,DNA Primers ,Glycoproteins ,Homeodomain Proteins ,Neuroectoderm ,Base Sequence ,Wnt signaling pathway ,Brain ,biology.organism_classification ,Blastula ,Molecular biology ,embryonic structures ,Homeobox ,Intercellular Signaling Peptides and Proteins ,Chordin ,Carrier Proteins ,Signal Transduction ,Developmental Biology - Abstract
The blastula Chordin- and Noggin-expressing (BCNE) center located in the dorsal animal region of the Xenopus blastula embryo contains both prospective anterior neuroectoderm and Spemann organizer precursor cells. Here we show that, contrary to previous reports, the canonical Wnt target homeobox genes, Double knockdown of these genes using antisense morpholinos in Xenopus laevis blocked head formation, reduced the expression of the other BCNE center genes, upregulated Bmp4 expression, and nullified hyperdorsalization by lithium chloride. Moreover, gain- and loss-of-function experiments showed that Siamois and Twin expression is repressed by the vegetal transcription factor VegT. We propose that VegT expression causes maternal beta-Catenin signals to restrict Siamois and Twin expression to the BCNE region. A two-step inhibition of BMP signals by Siamois and Twin-- first by transcriptional repression of Bmp4 and then by activation of the expression of the BMP inhibitors Chordin and Noggin--in the BCNE center is required for head formation.
- Published
- 2008
28. TAK1 determines susceptibility to endoplasmic reticulum stress and leptin resistance in the hypothalamus
- Author
-
Kazuhito, Sai, Sho, Morioka, Giichi, Takaesu, Nagendran, Muthusamy, H Troy, Ghashghaei, Hiroshi, Hanafusa, Kunihiro, Matsumoto, and Jun, Ninomiya-Tsuji
- Subjects
Leptin ,Mice, Knockout ,Sterol Regulatory Element Binding Proteins ,Mice ,Hypothalamus ,Animals ,Obesity ,Hyperphagia ,Endoplasmic Reticulum Stress ,MAP Kinase Kinase Kinases ,Dietary Fats ,Research Article - Abstract
Sustained endoplasmic reticulum (ER) stress disrupts normal cellular homeostasis and leads to the development of many types of human diseases, including metabolic disorders. TAK1 (also known as MAP3K7) is a member of the mitogen-activated protein kinase kinase kinase (MAP3K) family and is activated by a diverse set of inflammatory stimuli. Here, we demonstrate that TAK1 regulates ER stress and metabolic signaling through modulation of lipid biogenesis. We found that deletion of Tak1 increased ER volume and facilitated ER-stress tolerance in cultured cells, which was mediated by upregulation of sterol-regulatory-element-binding protein (SREBP)-dependent lipogenesis. In the in vivo setting, central nervous system (CNS)-specific Tak1 deletion upregulated SREBP-target lipogenic genes and blocked ER stress in the hypothalamus. Furthermore, CNS-specific Tak1 deletion prevented ER-stress-induced hypothalamic leptin resistance and hyperphagic obesity under a high-fat diet (HFD). Thus, TAK1 is a crucial regulator of ER stress in vivo, which could be a target for alleviation of ER stress and its associated disease conditions.
- Published
- 2015
29. C. elegans Tensin Promotes Axon Regeneration by Linking the Met-like SVH-2 and Integrin Signaling Pathways.
- Author
-
Naoki Hisamoto, Tatsuhiro Shimizu, Kazuma Asai, Yoshiki Sakai, Pastuhov, Strahil I., Hiroshi Hanafusa, and Kunihiro Matsumoto
- Subjects
AXONS ,MOTOR neurons ,MITOGEN-activated protein kinases ,PROTEIN-tyrosine kinases ,NERVOUS system regeneration - Abstract
Axon regeneration is a conserved mechanism induced by axon injury that initiates a neuronal response leading to regrowth of the axon. In Caenorhabditis elegans, the initiation of axon regeneration is regulated by the JNK MAP kinase (MAPK) pathway. We have previously identified a number of genes affecting the JNK pathway using an RNAi-based screen. Analysis of these genes, called the svh genes, has shed new light on the regulation of axon regeneration, revealing the involvement of a signaling cascade consisting of a growth factor SVH-1 and its receptor, the tyrosine kinase SVH-2. Here, we characterize the svh-6/tns-l gene, which is a homolog of mammalian tensin, and show that it is a positive regulator of axon regeneration in motor neurons. We demonstrate that TNS-1 interacts with tyrosine-autophosphorylated SVH-2 and the integrin J3 subunit PAT-3 via its SH2 and PTB domains, respectively, to promote axon regeneration. These results suggest that TNS-1 acts as an adaptor to link the SVH-2 and integrin signaling pathways. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
30. LRRK1 phosphorylation of Rab7 at S72 links trafficking of EGFR-containing endosomes to its effector RILP.
- Author
-
Hiroshi Hanafusa, Takuya Yagi, Haruka Ikeda, Naoki Hisamoto, Tomoki Nishioka, Kozo Kaibuchi, Kyoko Shirakabe, and Kunihiro Matsumoto
- Subjects
- *
EPIDERMAL growth factor receptors , *MOLECULAR motor proteins , *CELL receptors , *PHOSPHORYLATION - Abstract
Ligand-induced activation of epidermal growth factor receptor (EGFR) initiates trafficking events that re-localize the receptor from the cell surface to intracellular endocytic compartments. EGFR-containing endosomes are transported to lysosomes for degradation by the dynein–dynactin motor protein complex. However, this cargodependent endosomal trafficking mechanism remains largely uncharacterized. Here, we show that GTP-bound Rab7 is phosphorylated on S72 by leucine-rich repeat kinase 1 (LRRK1) at the endosomal membrane. This phosphorylation promotes the interaction of Rab7 (herein referring to Rab7a) with its effector RILP, resulting in recruitment of the dynein–dynactin complex to Rab7-positive vesicles. This, in turn, facilitates the dynein-driven transport of EGFR-containing endosomes toward the perinuclear region. These findings reveal a mechanism regulating the cargo-specific trafficking of endosomes. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
31. LRRK1 phosphorylation of Rab7 at Ser-72 links trafficking of EGFR-containing endosomes to its effector RILP.
- Author
-
Hiroshi Hanafusa, Takuya Yagi, Haruka Ikeda, Naoki Hisamoto, Tomoki Nishioka, Kozo Kaibuchi, Kyoko Shirakabe, and Kunihiro Matsumoto
- Subjects
EPIDERMAL growth factor receptors ,MOLECULAR motor proteins ,CELL receptors ,PHOSPHORYLATION - Abstract
Ligand-induced activation of epidermal growth factor receptor (EGFR) initiates trafficking events that re-localize the receptor from the cell surface to intracellular endocytic compartments. EGFR-containing endosomes are transported to lysosomes for degradation by the dynein-dynactin motor protein complex. However, this cargo-dependent endosomal trafficking mechanism remains largely uncharacterized. Here, we show that GTP-bound Rab7 is phosphorylated on Ser-72 by leucine-rich repeat kinase 1 (LRRK1) at the endosomal membrane. This phosphorylation promotes the interaction of Rab7 with its effector RILP, resulting in recruitment of the dynein-dynactin complex to Rab7-positive vesicles. This, in turn, facilitates the dynein-driven transport of EGFR-containing endosomes toward the perinuclear region. These findings reveal a mechanism regulating the cargo-specific trafficking of endosomes. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
32. Xenopus ILK (integrin-linked kinase) is required for morphogenetic movements during gastrulation
- Author
-
Hiroshi Hanafusa, Norihisa Masuyama, Morioh Kusakabe, Hiroaki Yamanaka, Eisuke Nishida, and Takayuki Yasunaga
- Subjects
Scaffold protein ,biology ,Integrin ,Embryogenesis ,Xenopus ,Cell Biology ,biology.organism_classification ,Cell biology ,Gastrulation ,embryonic structures ,Genetics ,biology.protein ,Integrin-linked kinase ,Signal transduction ,Axis elongation - Abstract
It has been suggested that ILK (integrin-linked kinase) participates in integrin- and growth factor-mediated signaling pathways and also functions as a scaffold protein at cell–extracellular matrix (ECM) adhesion sites. As the recently reported ILK knockout mice were found to die at the peri-implantation stage, the stage specific to mammals, little is known about the function of ILK in early developmental processes common to every vertebrate. To address this, we isolated a Xenopus ortholog of ILK (XeILK) and characterized its role in early Xenopus embryogenesis. XeILK was expressed constitutively and ubiquitously throughout the early embryogenesis. Depletion of XeILK with morpholino oligonucleotides (XeILK MO) caused severe defects in blastopore closure and axis elongation without affecting the mesodermal specification. Furthermore, XeILK MO was found to interfere with cell–cell and cell–ECM adhesions in dorsal marginal zone explants and to result in a significant loss of cell–ECM adhesions in activin-treated dissociated animal cap cells. These results thus indicate that XeILK plays an essential role in morphogenetic movements during gastrulation.
- Published
- 2005
33. Shp2, an SH2-containing Protein-tyrosine Phosphatase, Positively Regulates Receptor Tyrosine Kinase Signaling by Dephosphorylating and Inactivating the Inhibitor Sprouty
- Author
-
Hiroshi Hanafusa, Kunihiro Matsumoto, Satoru Torii, Eisuke Nishida, and Takayuki Yasunaga
- Subjects
SH2 Domain-Containing Protein Tyrosine Phosphatases ,Protein Tyrosine Phosphatase, Non-Receptor Type 11 ,Protein tyrosine phosphatase ,Biochemistry ,Receptor tyrosine kinase ,src Homology Domains ,Mice ,chemistry.chemical_compound ,Animals ,Phosphorylation ,Molecular Biology ,Binding Sites ,biology ,Chemistry ,Cell Membrane ,fungi ,Intracellular Signaling Peptides and Proteins ,Receptor Protein-Tyrosine Kinases ,Tyrosine phosphorylation ,Cell Biology ,Cell biology ,Enzyme Activation ,Protein Transport ,Mitogen-activated protein kinase ,biology.protein ,Cancer research ,GRB2 ,Protein Tyrosine Phosphatases ,Signal transduction ,Signal Transduction ,Proto-oncogene tyrosine-protein kinase Src - Abstract
Src homology 2-containing phosphotyrosine phosphatase (Shp2) functions as a positive effector in receptor tyrosine kinase (RTK) signaling immediately proximal to activated receptors. However, neither its physiological substrate(s) nor its mechanism of action in RTK signaling has been defined. In this study, we demonstrate that Sprouty (Spry) is a possible target of Shp2. Spry acts as a conserved inhibitor of RTK signaling, and tyrosine phosphorylation of Spry is indispensable for its inhibitory activity. Shp2 was able to dephosphorylate fibroblast growth factor receptor-induced phosphotyrosines on Spry both in vivo and in vitro. Shp2-mediated dephosphorylation of Spry resulted in dissociation of Spry from Grb2. Furthermore, Shp2 could reverse the inhibitory effect of Spry on FGF-induced neurite outgrowth and MAP kinase activation. These findings suggest that Shp2 acts as a positive regulator in RTK signaling by dephosphorylating and inactivating Spry.
- Published
- 2004
34. Phosphorylation of CLIP-170 by LRRK1 regulates EGFR trafficking by promoting recruitment of p150Glued to MT plus-ends
- Author
-
Takashi Watanabe, Strahil Iv. Pastuhov, Hiroshi Hanafusa, Kunihiro Matsumoto, Kozo Kaibuchi, Tomoki Nishioka, and Shin Kedashiro
- Subjects
Gene product ,Microtubule ,Endosome ,Protein subunit ,Endocytic cycle ,Dynactin ,Phosphorylation ,Cell Biology ,Biology ,Kinase activity ,Cell biology - Abstract
Ligand-induced activation of the EGF receptor (EGFR) initiates trafficking events that relocalize the receptors from the cell surface to intracellular endocytic compartments. Early endosomes containing activated EGFR migrate along microtubules as they mature into late endosomes. We have recently shown that LRRK1, which is related to the familial Parkinsonism gene product Park8/LRRK2, regulates this EGFR transport in a manner dependent on LRRK1 kinase activity. However, the downstream targets of LRRK1 that may modulate this transport function have not been identified. Here, we identify CLIP-170, a microtubule plus-end protein, as a substrate of LRRK1. LRRK1 phosphorylates CLIP-170 at Thr-1384, located in its C-terminal zinc knuckle motif, and this promotes its association with dynactin–dynein complexes. We find that LRRK1 phosphorylation of CLIP-170 causes the accumulation of p150Glued, a subunit of dynactin, at microtubule plus-ends, thereby facilitating the migration of EGFR-containing endosomes. Thus, our findings provide new mechanistic insights into the dynein-driven transport of EGFR.
- Published
- 2014
35. The TGF‐β family member derrière is involved in regulation of the establishment of left–right asymmetry
- Author
-
Norihisa Masuyama, Hiroshi Hanafusa, Hiroshi Shibuya, Eisuke Nishida, and Morioh Kusakabe
- Subjects
Embryo, Nonmammalian ,Microinjections ,Xenopus ,media_common.quotation_subject ,Blotting, Western ,Molecular Sequence Data ,Embryonic Development ,Xenopus Proteins ,Biology ,Biochemistry ,Asymmetry ,Transforming Growth Factor beta ,Genetics ,Animals ,RNA, Messenger ,Protein Precursors ,Growth Substances ,Molecular Biology ,In Situ Hybridization ,Body Patterning ,Glycoproteins ,media_common ,Scientific Reports ,Intracellular Signaling Peptides and Proteins ,Heart ,Zebrafish Proteins ,Molecular pathway ,biology.organism_classification ,Immunohistochemistry ,Cell biology ,Viscera ,Family member ,Oocytes ,Intercellular Signaling Peptides and Proteins ,Female ,Ectopic expression ,NODAL ,Transforming growth factor - Abstract
Although a number of genes that are involved in the establishment of left-right asymmetry have been identified, earlier events in the molecular pathway developing left-right asymmetry remain to be elucidated. Here we present evidence suggesting that the transforming growth factor-beta family member derrière is involved in the development of left-right asymmetry in Xenopus embryos. Ectopic expression of derrière on the right side can fully invert cardiac and visceral left-right orientation and nodal expression, and expression of a dominant-negative form of derrière on the left side can partially randomize the left-right orientation and nodal expression. Moreover, while expression of the dominant-negative derrière does not inhibit the activity of Vg1 directly, it can rescue the altered left-right orientation induced by Vg1. Vg1 can induce derrière in animal cap explants. These results suggest that derrière is involved in earlier molecular pathways developing the left-right asymmetry.
- Published
- 2000
36. Improvement of the spin-on-glass process by ion implantation for highly reliable MOS devices
- Author
-
Kaori Misawa, Makoto Akizuki, Hiroyuki Aoe, Kazunobu Mameno, Hiroyuki Watanabe, Hideki Mizuhara, K. Yodoshi, Hiroshi Hanafusa, and Mamoru Arimoto
- Subjects
Materials science ,Hydrogen ,business.industry ,Analytical chemistry ,General Physics and Astronomy ,chemistry.chemical_element ,Surfaces and Interfaces ,General Chemistry ,Dielectric ,Condensed Matter Physics ,Surfaces, Coatings and Films ,Ion ,Stress (mechanics) ,Ion implantation ,chemistry ,Optoelectronics ,Wafer ,Field-effect transistor ,Diffusion (business) ,business - Abstract
We have developed a novel process for intermetal dielectrics (IMD) using ion-implanted organic spin-on glass (SOG). Ion implantation into the SOG films improves the hot carrier (HC) reliability, which deteriorates from the use of organic SOG films. The content of mobile water-related species in the organic SOG films, which was examined by thermal desorption spectrometry (TDS), is reduced by Ar+-ion implantation under the conditions of 140 keV, 1 × 1015 ions/cm2; while on the contrary, the content of mobile hydrogen in SOG is increased by ion implantation. The change in the mechanical stress due to ion implantation is small, because ion implantation shrinks SOG only in the direction vertical to the wafer surface. Thus the change in stress due to ion implantation scarcely affect the electrical characteristics of the underlying MOS-FET. Therefore the deterioration of HC reliability for the MOS devices with SOG as IMD is well explained by a water diffusion model rather than by a hydrogen diffusion and mechanical stress model.
- Published
- 1997
37. [Regulation of intracellular trafficking of the EGF receptor by ROCO family kinase LRRK1]
- Author
-
Hiroshi, Hanafusa and Kunihiro, Matsumoto
- Subjects
ErbB Receptors ,Protein Transport ,Cell Transformation, Neoplastic ,Humans ,Endosomes ,Protein Serine-Threonine Kinases ,Lysosomes ,GRB2 Adaptor Protein - Published
- 2012
38. EGFR-dependent phosphorylation of leucine-rich repeat kinase LRRK1 is important for proper endosomal trafficking of EGFR
- Author
-
Hiroshi Hanafusa, Atsuki Nara, Kunihiro Matsumoto, and Kouki Ishikawa
- Subjects
Endosome ,Movement ,Down-Regulation ,Endosomes ,Biology ,Leucine-rich repeat ,Protein Serine-Threonine Kinases ,Endocytosis ,Epidermal growth factor ,Humans ,Kinase activity ,Phosphorylation ,Molecular Biology ,Sequence Deletion ,Feedback, Physiological ,Binding Sites ,Epidermal Growth Factor ,Kinase ,Escherichia coli Proteins ,Cell Biology ,Articles ,Cell biology ,Enzyme Activation ,ErbB Receptors ,Protein Transport ,Amino Acid Substitution ,Mutagenesis ,Membrane Trafficking ,Cyclin-dependent kinase 8 ,RNA ,Tyrosine ,Guanosine Triphosphate ,Bacterial Outer Membrane Proteins ,HeLa Cells - Abstract
Endocytosis and subsequent delivery of activated EGFR to lysosomes are essential for the termination of EGFR signaling. It is shown that EGFR regulates the kinase activity of LRRK1 via tyrosine phosphorylation and that this is required for proper regulation of endosomal trafficking of EGFR., Ligand-induced activation of the epidermal growth factor receptor (EGFR) initiates trafficking events that relocalize the receptors from the cell surface to intracellular endocytic compartments. We recently reported that leucine-rich repeat kinase 1 (LRRK1) is involved in the trafficking of EGFR from early to late endosomes. In this study, we demonstrate that EGFR regulates the kinase activity of LRRK1 via tyrosine phosphorylation and that this is required for proper endosomal trafficking of EGFR. Phosphorylation of LRRK1 at Tyr-944 results in reduced LRRK1 kinase activity. Mutation of LRRK1 Tyr-944 (Y944F) abolishes EGF-stimulated tyrosine phosphorylation, resulting in hyperactivation of LRRK1 kinase activity and enhanced motility of EGF-containing endosomes toward the perinuclear region. The compartments in which EGFR accumulates are mixed endosomes and are defective in the proper formation of intraluminal vesicles of multivesicular bodies. These results suggest that feedback down-regulation of LRRK1 kinase activity by EGFR plays an important role in the appropriate endosomal trafficking of EGFR.
- Published
- 2012
39. Leucine-rich repeat kinase LRRK1 regulates endosomal trafficking of the EGF receptor
- Author
-
Shun-ichiro Iemura, Atsuki Nara, Tsukasa Saigo, Tohru Natsume, Hiroshi Hanafusa, Hiroshi Shibuya, Shin Kedashiro, Masayuki Komada, Kouki Ishikawa, and Kunihiro Matsumoto
- Subjects
MAPK/ERK pathway ,Multidisciplinary ,biology ,Endosome ,Endocytic cycle ,General Physics and Astronomy ,General Chemistry ,General Biochemistry, Genetics and Molecular Biology ,Article ,Cell biology ,Protein kinase domain ,Epidermal growth factor ,biology.protein ,Epidermal growth factor receptor ,GRB2 ,Kinase activity - Abstract
Activation of the epidermal growth factor receptor (EGFR) not only initiates multiple signal-transduction pathways, including the MAP kinase (MAPK) pathway, but also triggers trafficking events that relocalize receptors from the cell surface to intracellular endocytic compartments. In this paper, we demonstrate that leucine-rich repeat kinase LRRK1, which contains a MAPKKK-like kinase domain, forms a complex with activated EGFR through an interaction with Grb2. Subsequently, LRRK1 and epidermal growth factor (EGF) are internalized and co-localized in early endosomes. LRRK1 regulates EGFR transport from early to late endosomes and regulates the motility of EGF-containing early endosomes in a manner dependent on its kinase activity. Furthermore, LRRK1 serves as a scaffold facilitating the interaction of EGFR with the endosomal sorting complex required for transport-0 complex, thus enabling efficient sorting of EGFR to the inner vesicles of multivesicular bodies. Our findings provide the first evidence that a MAPKKK-like protein regulates the endosomal trafficking of EGFR., Activation of the epidermal growth factor receptor can result in its internalization and subsequent intracellular trafficking. In this study, the authors show that leucine-rich repeat kinase-1 can bind to the receptor and regulate its trafficking between different endosomal compartments.
- Published
- 2011
40. Development of a new heterojunction structure (ACJ -HIT) and its application to polycrystalline silicon solar cells
- Author
-
Toru Sawada, Shoichi Nakano, Yukinori Kuwano, Shinya Tsuda, Mikio Taguchi, Takao Matsuyama, Tsuysohi Takahama, Akio Takeoka, Shigeru Kuroda, Makoto Tanaka, and Hiroshi Hanafusa
- Subjects
Materials science ,Passivation ,Renewable Energy, Sustainability and the Environment ,business.industry ,Energy conversion efficiency ,Heterojunction ,Plasma ,engineering.material ,Condensed Matter Physics ,Polymer solar cell ,Electronic, Optical and Magnetic Materials ,law.invention ,Monocrystalline silicon ,Polycrystalline silicon ,law ,Solar cell ,engineering ,Optoelectronics ,Electrical and Electronic Engineering ,business - Abstract
A new solar cell structure named HIT (Heterojunction with Intrinsic Thin layer) has been developed based on new artificially constructed junction (ACJ) technology. In this structure a non-doped a-Si thin layer was inserted between the p(a-Si)/n(c-Si) heterojunction, improving the output characteristics and achieving a conversion efficiency of 18.1%. This structure was applied to cast polycrystalline silicon solar cells of a practical size. A high conversion efficeincy of 13.6% was obtained with a cell size of 10 cm × 10 cm using various technologies, including hydrogen plasma passivation.
- Published
- 1993
41. Preparation of poly-si by excimer laser annealing with solidification process control
- Author
-
Hiroyuki Kuriyama, Shoichi Nakano, Seiichi Kiyama, Noguchi Shigeru, Hiroshi Hanafusa, and Takashi Kuwahara
- Subjects
Materials science ,Excimer laser ,Computer Networks and Communications ,business.industry ,medicine.medical_treatment ,General Physics and Astronomy ,Substrate (electronics) ,engineering.material ,Laser ,Grain size ,Amorphous solid ,law.invention ,Polycrystalline silicon ,law ,Thin-film transistor ,medicine ,engineering ,Optoelectronics ,Irradiation ,Electrical and Electronic Engineering ,business - Abstract
Polycrystalline silicon (poly-Si) recrystallized by a conventional excimer laser has a small grain of about a few hundred A and a mobility of about 100 cm2/Vs at most. It is believed that such small grain size could be due to a fast solidification velocity based on the authors's study of the solidification control of molten Si. A thermal analysis by computer simulation indicates that the solidification velocity can be lowered by heating substrate (400°C) during laser irradiation and thinning an amorphous Si (a-Si) film (500 A). Poly-Si with a large grain (5000 A) and high mobility (280 cm2/Vs) has been fabricated by excimer laser annealing in the present study. Nonuniformity of characteristics of poly-Si in an overlapping region of the laser spot also is examined. An uncompleted melting of a mixed region of fine poly-Si and a-Si generated at an excimer laser spot edge is the cause for such nonuniformity. An overlapping region of a laser spot is found to become very uniform by using the present solidification control method. Nonuniformity of mobility is improved from ±20 to ±8 percent in the present study. This method is a very useful low-temperature processing for fabricating large-sized poly-Si films.
- Published
- 1993
42. Regulation of ERK activity duration by Sprouty contributes to dorsoventral patterning
- Author
-
Kunihiro Matsumoto, Eisuke Nishida, and Hiroshi Hanafusa
- Subjects
MAPK/ERK pathway ,Embryo, Nonmammalian ,Xenopus ,Embryonic Development ,Context (language use) ,Biology ,Cell fate determination ,Xenopus Proteins ,Mesoderm ,Xenopus laevis ,Animals ,Protein Isoforms ,Cell Lineage ,Extracellular Signal-Regulated MAP Kinases ,Adaptor Proteins, Signal Transducing ,Body Patterning ,Regulation of gene expression ,Intracellular Signaling Peptides and Proteins ,Signal transducing adaptor protein ,Gene Expression Regulation, Developmental ,Membrane Proteins ,Embryo ,Cell Differentiation ,Cell Biology ,biology.organism_classification ,Phosphoproteins ,Cell biology ,Enzyme Activation ,Mesoderm formation ,Signal Transduction - Abstract
Distinct modes of ERK activation, sustained or transient, are essential for cell fate decision in cultured cells. Here we show that Xenopus laevis Sprouty2 (XSpry2) controls the duration of ERK activity and thereby contributes to the establishment of dorsoventral patterning during mesoderm formation. Furthermore, Xenopus Fos (XFos) can function as a molecular sensor of the ERK signalling duration in Xenopus embryos. This work provides the first evidence that regulating the duration of ERK activity contributes to cell fate decisions in the context of the whole organism.
- Published
- 2008
43. LRRK1-phosphorylated CLIP-170 regulates EGFR trafficking by recruiting p150Glued to microtubule plus ends
- Author
-
Hiroshi Hanafusa, Strahil Iv. Pastuhov, Kozo Kaibuchi, Tomoki Nishioka, Shin Kedashiro, Kunihiro Matsumoto, and Takashi Watanabe
- Subjects
Endosome ,Endosomes ,Protein Serine-Threonine Kinases ,Biology ,Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 ,Bioinformatics ,Endocytosis ,Microtubules ,Microtubule ,Humans ,Epidermal growth factor receptor ,Phosphorylation ,Kinase activity ,Dyneins ,Dynactin Complex ,Cell Biology ,Neoplasm Proteins ,Cell biology ,ErbB Receptors ,DCTN1 ,biology.protein ,Dynactin ,Microtubule-Associated Proteins ,HeLa Cells ,Protein Binding - Abstract
The binding of ligand to epidermal growth factor receptor (EGFR) causes the receptor to become activated and stimulates the endocytosis of EGFR. Early endosomes containing activated EGFR migrate along microtubules as they mature into late endosomes. We have recently shown that LRRK1, which is related to the familial Parkinsonism gene product Park8 (also known as LRRK2), regulates this EGFR transport in a manner dependent on LRRK1 kinase activity. However, the downstream targets of LRRK1 that might modulate this transport function have not been identified. Here, we identify CLIP-170 (also known as CLIP1), a microtubule plus-end protein, as a substrate of LRRK1. LRRK1 phosphorylates CLIP-170 at Thr1384, located in its C-terminal zinc knuckle motif, and this promotes the association of CLIP-170 with dynein–dynactin complexes. We find that LRRK1-mediated phosphorylation of CLIP-170 causes the accumulation of p150 Glued (also known as DCTN1) a subunit of dynactin, at microtubule plus ends, thereby facilitating the migration of EGFR-containing endosomes. Thus, our findings provide new mechanistic insights into the dynein-driven transport of EGFR.
- Published
- 2015
44. Xenopus ILK (integrin-linked kinase) is required for morphogenetic movements during gastrulation
- Author
-
Takayuki, Yasunaga, Morioh, Kusakabe, Hiroaki, Yamanaka, Hiroshi, Hanafusa, Norihisa, Masuyama, and Eisuke, Nishida
- Subjects
Xenopus laevis ,Molecular Sequence Data ,Cell Adhesion ,Animals ,Amino Acid Sequence ,Gastrula ,Protein Serine-Threonine Kinases ,Xenopus Proteins ,Extracellular Matrix - Abstract
It has been suggested that ILK (integrin-linked kinase) participates in integrin- and growth factor-mediated signaling pathways and also functions as a scaffold protein at cell-extracellular matrix (ECM) adhesion sites. As the recently reported ILK knockout mice were found to die at the peri-implantation stage, the stage specific to mammals, little is known about the function of ILK in early developmental processes common to every vertebrate. To address this, we isolated a Xenopus ortholog of ILK (XeILK) and characterized its role in early Xenopus embryogenesis. XeILK was expressed constitutively and ubiquitously throughout the early embryogenesis. Depletion of XeILK with morpholino oligonucleotides (XeILK MO) caused severe defects in blastopore closure and axis elongation without affecting the mesodermal specification. Furthermore, XeILK MO was found to interfere with cell-cell and cell-ECM adhesions in dorsal marginal zone explants and to result in a significant loss of cell-ECM adhesions in activin-treated dissociated animal cap cells. These results thus indicate that XeILK plays an essential role in morphogenetic movements during gastrulation.
- Published
- 2005
45. Sprouty1 and Sprouty2 provide a control mechanism for the Ras/MAPK signalling pathway
- Author
-
Hiroshi Hanafusa, Eisuke Nishida, Takayuki Yasunaga, and Satoru Torii
- Subjects
MAPK/ERK pathway ,Time Factors ,Xenopus ,Protein Tyrosine Phosphatase, Non-Receptor Type 11 ,Protein tyrosine phosphatase ,PC12 Cells ,Receptor tyrosine kinase ,Mice ,Phosphorylation ,Luciferases ,Genes, Dominant ,Neurons ,biology ,Intracellular Signaling Peptides and Proteins ,Cell biology ,Protein Transport ,Fibroblast growth factor receptor ,COS Cells ,GRB2 ,biological phenomena, cell phenomena, and immunity ,Mitogen-Activated Protein Kinases ,Dimerization ,Platelet-derived growth factor receptor ,Plasmids ,Protein Binding ,DNA, Complementary ,MAP Kinase Signaling System ,Immunoblotting ,Molecular Sequence Data ,Nerve Tissue Proteins ,Binding, Competitive ,Cell Line ,Growth factor receptor ,Animals ,Humans ,Amino Acid Sequence ,Adaptor Proteins, Signal Transducing ,Sequence Homology, Amino Acid ,Cell Membrane ,Membrane Proteins ,Cell Biology ,Phosphoproteins ,Precipitin Tests ,Receptors, Fibroblast Growth Factor ,Rats ,Enzyme Activation ,Microscopy, Fluorescence ,SPRY2 ,Mutation ,biology.protein ,ras Proteins ,Tyrosine ,Cattle ,Protein Tyrosine Phosphatases ,Peptides ,Protein Processing, Post-Translational ,HeLa Cells - Abstract
Sprouty (Spry) inhibits signalling by receptor tyrosine kinases; however, the molecular mechanism underlying this function has not been defined. Here we show that after stimulation by growth factors Spry1 and Spry2 translocate to the plasma membrane and become phosphorylated on a conserved tyrosine. Next, they bind to the adaptor protein Grb2 and inhibit the recruitment of the Grb2–Sos complex either to the fibroblast growth factor receptor (FGFR) docking adaptor protein FRS2 or to Shp2. Membrane translocation of Spry is necessary for its phosphorylation, which is essential for its inhibitor activity. A tyrosine-phosphorylated octapeptide derived from mouse Spry2 inhibits Grb2 from binding FRS2, Shp2 or mouse Spry2 in vitro and blocks activation of the extracellular-signal-regulated kinase (ERK) in cells stimulated by growth factor. A non-phosphorylated Spry mutant cannot bind Grb2 and acts as a dominant negative, inducing prolonged activation of ERK in response to FGF and promoting the FGF-induced outgrowth of neurites in PC12 cells. Our findings suggest that Spry functions in a negative feedback mechanism in which its inhibitor activity is controlled rapidly and reversibly by post-translational mechanisms.
- Published
- 2002
46. Xenopus FRS2 is involved in early embryogenesis in cooperation with the Src family kinase Laloo
- Author
-
Eisuke Nishida, Hiroshi Hanafusa, Norihisa Masuyama, and Morioh Kusakabe
- Subjects
Embryo, Nonmammalian ,Macromolecular Substances ,Molecular Sequence Data ,Xenopus ,Embryonic Development ,Xenopus Proteins ,Fibroblast growth factor ,Biochemistry ,FGF and mesoderm formation ,chemistry.chemical_compound ,Xenopus laevis ,Genetics ,Animals ,Humans ,Src family kinase ,Amino Acid Sequence ,Cloning, Molecular ,Phosphorylation ,Molecular Biology ,In Situ Hybridization ,Adaptor Proteins, Signal Transducing ,Body Patterning ,biology ,Scientific Reports ,Membrane Proteins ,Tyrosine phosphorylation ,biology.organism_classification ,Phosphoproteins ,Molecular biology ,Receptors, Fibroblast Growth Factor ,Cell biology ,Fibroblast Growth Factors ,src-Family Kinases ,chemistry ,Mesoderm formation ,biology.protein ,GRB2 ,Sequence Alignment ,Protein Binding ,Signal Transduction - Abstract
FRS2 has been identified in mammalian cells as a protein that is tyrosine phosphorylated and binds to Grb2 and Shp2 in response to fibroblast growth factor (FGF) or nerve growth factor (NGF) stimulation. But neither its existence in other vertebrate classes or invertebrates nor its function during embryonic development has been defined. Here we have identified and characterized a Xenopus homolog of FRS2 (xFRS2). xFRS2 is tyrosine phosphorylated in early embryos, and overexpression of an unphosphorylatable form of xFRS2 interferes with FGF-dependent mesoderm formation. The Src family kinase Laloo, which was shown to function in FGF signaling during early Xenopus development, binds to xFRS2 and promotes tyrosine phosphorylation of xFRS2. Moreover, xFRS2 and Laloo are shown to bind to Xenopus FGF receptor 1. These results suggest that xFRS2 plays an important role in FGF signaling in cooperation with Laloo during embryonic development.
- Published
- 2001
47. Involvement of the p38 mitogen-activated protein kinase pathway in transforming growth factor-beta-induced gene expression
- Author
-
Hiroshi Hanafusa, Eisuke Nishida, Norihisa Masuyama, Hiroshi Shibuya, Kunihiro Matsumoto, Michiru Nishita, Jun Ninomiya-Tsuji, and Jun-ichi Fujisawa
- Subjects
MAP Kinase Kinase 6 ,Mitogen-activated protein kinase kinase ,Biochemistry ,p38 Mitogen-Activated Protein Kinases ,MAP2K7 ,Transforming Growth Factor beta ,Animals ,ASK1 ,Genes, Tumor Suppressor ,c-Raf ,Cyclic AMP Response Element-Binding Protein ,Molecular Biology ,Cells, Cultured ,Leucine Zippers ,biology ,MAP kinase kinase kinase ,Activating Transcription Factor 2 ,Chemistry ,Cyclin-dependent kinase 4 ,Cyclin-dependent kinase 2 ,Cell Biology ,MAP Kinase Kinase Kinases ,Molecular biology ,DNA-Binding Proteins ,Gene Expression Regulation ,Calcium-Calmodulin-Dependent Protein Kinases ,biology.protein ,Trans-Activators ,Cyclin-dependent kinase 9 ,Mitogen-Activated Protein Kinases ,Protein Kinases ,Signal Transduction ,Transcription Factors - Abstract
Transforming growth factor-beta (TGF-beta)-activated kinase 1 (TAK1), a member of the mitogen-activated protein kinase kinase kinase family, is suggested to be involved in TGF-beta-induced gene expression, but the signaling mechanism from TAK1 to the nucleus remains largely undefined. We have found that p38 mitogen-activated protein kinase, and its direct activator MKK6 are rapidly activated in response to TGF-beta. Expression of dominant negative MKK6 or dominant negative TAK1 inhibited the TGF-beta-induced transcriptional activation as well as the p38 activation. Constitutive activation of the p38 pathway in the absence of TGF-beta induced the transcriptional activation, which was enhanced synergistically by coexpression of Smad2 and Smad4 and was inhibited by expression of the C-terminal truncated, dominant negative Smad4. Furthermore, we have found that activating transcription factor-2 (ATF-2), which is known as a nuclear target of p38, becomes phosphorylated in the N-terminal activation domain in response to TGF-beta, that ATF-2 forms a complex with Smad4, and that the complex formation is enhanced by TGF-beta. In addition, expression of a nonphosphorylatable form of ATF-2 inhibited the TGF-beta-induced transcriptional activation. These results show that the p38 pathway is activated by TGF-beta and is involved in the TGF-beta-induced transcriptional activation by regulating the Smad-mediated pathway.
- Published
- 1999
48. A novel SAPK/JNK kinase, MKK7, stimulated by TNFalpha and cellular stresses
- Author
-
Hiroshi Hanafusa, Norihisa Masuyama, Tetsuo Moriguchi, Eisuke Nishida, Fumiko Toyoshima, and Yukiko Gotoh
- Subjects
MAPK/ERK pathway ,DNA, Complementary ,Transcription, Genetic ,MAP Kinase Kinase 4 ,p38 mitogen-activated protein kinases ,Molecular Sequence Data ,MAP Kinase Kinase 7 ,Biology ,Mitogen-activated protein kinase kinase ,Protein Serine-Threonine Kinases ,General Biochemistry, Genetics and Molecular Biology ,MAP2K7 ,Substrate Specificity ,Mice ,Animals ,Amino Acid Sequence ,Cloning, Molecular ,Protein kinase A ,Molecular Biology ,Mitogen-Activated Protein Kinase Kinases ,General Immunology and Microbiology ,MAP kinase kinase kinase ,Sequence Homology, Amino Acid ,Kinase ,Tumor Necrosis Factor-alpha ,General Neuroscience ,JNK Mitogen-Activated Protein Kinases ,Protein-Tyrosine Kinases ,Molecular biology ,Cell biology ,Enzyme Activation ,Transcription Factor AP-1 ,Mitogen-activated protein kinase ,Calcium-Calmodulin-Dependent Protein Kinases ,biology.protein ,Mitogen-Activated Protein Kinases ,Protein Kinases ,Signal Transduction ,Research Article - Abstract
Stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK), a member of the MAP kinase (MAPK) superfamily, is thought to play a key role in a variety of cellular responses. To date, SEK1/MKK4, one of the MAP kinase kinase (MAPKK) family of molecules, is the only SAPK/JNK kinase that has been cloned. Here we have cloned, identified and characterized a novel member of the mammalian MAPKKs, designated MKK7. MKK7 is most similar to the mediator of morphogenesis, hemipterous (hep), in Drosophila. Immunochemical studies have identified MKK7 as one of the major SAPK/JNK-activating kinases in osmotically shocked cells. While SEK1/MKK4 can activate both the SAPK/JNK and p38 subgroups of the MAPK superfamily, MKK7 is specific for the SAPK/JNK subgroup. MKK7 is activated strongly by tumour necrosis factor alpha (TNFalpha) as well as by environmental stresses, whereas SEK1/MKK4 is not activated by TNFalpha. Column fractionation studies have shown that MKK7 is a major activator for SAPK/JNK in the TNFalpha-stimulated pathway. Moreover, we have found that overexpression of MKK7 enhances transcription from an AP-1-dependent reporter construct. Thus, MKK7 is an evolutionarily conserved MAPKK isoform which is specific for SAPK/JNK, is involved in AP-1-dependent transcription and may be a crucial mediator of TNFalpha signalling.
- Published
- 1998
49. TAK1 determines susceptibility to endoplasmic reticulum stress and leptin resistance in the hypothalamus.
- Author
-
Kazuhito Sai, Sho Morioka, Giichi Takaesu, Muthusamy, Nagendran, Ghashghaei, H. Troy, Hiroshi Hanafusa, Kunihiro Matsumoto, and Jun Ninomiya-Tsuji
- Subjects
ENDOPLASMIC reticulum ,ORGANELLES ,HYPOTHALAMUS ,OBESITY ,METABOLIC disorders - Abstract
Sustained endoplasmic reticulum (ER) stress disrupts normal cellular homeostasis and leads to the development of many types of human diseases, including metabolic disorders. TAK1 (also known as MAP3K7) is a member of the mitogen-activated protein kinase kinase kinase (MAP3K) family and is activated by a diverse set of inflammatory stimuli. Here, we demonstrate that TAK1 regulates ER stress and metabolic signaling through modulation of lipid biogenesis. We found that deletion of Tak1 increased ER volume and facilitated ER-stress tolerance in cultured cells, which was mediated by upregulation of sterol-regulatory-element-binding protein (SREBP)- dependent lipogenesis. In the in vivo setting, central nervous system (CNS)-specific Tak1 deletion upregulated SREBP-target lipogenic genes and blocked ER stress in the hypothalamus. Furthermore, CNS-specific Tak1 deletion prevented ER-stress-induced hypothalamic leptin resistance and hyperphagic obesity under a high-fat diet (HFD). Thus, TAK1 is a crucial regulator of ER stress in vivo, which could be a target for alleviation of ER stress and its associated disease conditions. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
50. A fast search method of similar strings from dictionaries
- Author
-
Hiroshi Hanafusa, Nobuo Fujisawa, Kazuhiro Morita, Jun-ichi Aoe, El-Sayed Atlam, and Masao Fuketa
- Subjects
TheoryofComputation_MISCELLANEOUS ,Information retrieval ,Similarity (geometry) ,String-to-string correction problem ,Computer Networks and Communications ,String (computer science) ,Industrial and Manufacturing Engineering ,Substring ,Computer Science Applications ,n-gram ,String kernel ,Edit distance ,Jaro–Winkler distance ,Electrical and Electronic Engineering ,Software ,Information Systems ,Mathematics - Abstract
The World Wide Web is growing ever more rapidly, and there are benefits from rich information. Moreover, demands for retrieving similar strings to an input string from dictionaries have been increasing. The edit distance is necessary to retrieve information from a large amount of data using the similarity between two strings. However, drawback of this method is time consumption because the input string must be compared with all strings in dictionaries. This study proposes a new technique for retrieving similar strings from dictionaries at high speed. The method presented can retrieve all similar strings 14 times faster than unigram methods although the edit distance is 3.
- Published
- 2011
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.