A5054997151- Cellular transport and secretion
- Retinal Development and Disorders
- Muscle Physiology and Disorders
- Endoplasmic Reticulum Stress and Disease
- Autophagy in Disease and Therapy
- Alzheimer's disease research and treatments
- Biotin and Related Studies
- Cell death mechanisms and regulation
- Ubiquitin and proteasome pathways
- Calcium signaling and nucleotide metabolism
- Lipid Membrane Structure and Behavior
- Nutrition and Health in Aging
- Pain Mechanisms and Treatments
- Fungal and yeast genetics research
- Monoclonal and Polyclonal Antibodies Research
- Lysosomal Storage Disorders Research
- RNA Interference and Gene Delivery
- TGF-β signaling in diseases
- Biofuel production and bioconversion
- Neurobiology and Insect Physiology Research
- Cell Adhesion Molecules Research
- Toxoplasma gondii Research Studies
- Fibromyalgia and Chronic Fatigue Syndrome Research
- Advanced Radiotherapy Techniques
- GDF15 and Related Biomarkers
Nagoya University
2018-2024
Fukushima Medical University
2007-2018
National Institutes of Health
2005-2007
Health and Human Development (2HD) Research Network
2006
Osaka University
1998-2005
Eunice Kennedy Shriver National Institute of Child Health and Human Development
2005
National Institute for Basic Biology
1998
Tokyo University of the Arts
1996
The University of Tokyo
1996
Cathepsin D-deficient (CD-/-) mice have been shown to manifest seizures and become blind near the terminal stage [approximately postnatal day (P) 26]. We therefore examined morphological, immunocytochemical, biochemical features of CNS tissues these mice. By electron microscopy, autophagosome/autolysosome-like bodies containing part cytoplasm, granular osmiophilic deposits, fingerprint profiles were demonstrated in neuronal perikarya CD-/- mouse brains after P20. Autophagosomes deposits...
Mutation in the Saccharomyces cerevisiae APG14 gene causes a defect autophagy. Cloning and structural analysis of revealed that encodes novel hydrophilic protein with predicted molecular mass 40.5 kDa, Apg14p has coiled-coil motif at its N terminus region. We found overproduction partially reversed autophagy induced by apg6-1 mutation. The mutant was to be defective not only but also sorting carboxypeptidase Y (CPY), vacuolar-soluble hydrolase, vacuole. However, overexpression did alter CPY...
The retromer is a cytosolic/peripheral membrane protein complex that mediates the retrieval of cation-independent mannose 6-phosphate receptor from endosomes to trans-Golgi network (TGN) in mammalian cells. Previous studies showed comprises three proteins, named Vps26, Vps29, and Vps35, plus sorting nexin, SNX1. There conflicting evidence, however, as whether homologous SNX2, truly component retromer. In addition, nature subunit interactions assembly are poorly understood. We have addressed...
Recent findings have suggested that the autophagic isolation membrane (IM) might originate from a domain of endoplasmic reticulum (ER) called omegasome. However, morphological relationships between ER, omegasome, and IM remain unclear. In present study, we found hybrid structures composed double FYVE domain-containing protein 1 (DFCP1)-positive omegasome accumulated in Atg3-deficient mouse embryonic fibroblasts (MEFs). Moreover, correlative light electron microscopy immunoelectron revealed...
The ubiquitin-proteasome system and autophagy are crucially important for proteostasis in cells. These pathways interdependent, dysfunction either pathway causes accumulation of ubiquitin-positive aggregates, a hallmark human pathological conditions. To elucidate vivo compensatory action(s) against proteasomal dysfunction, we developed mice with reduced proteasome activity their livers. mutant exhibited severe liver damage, accompanied by formation aggregates positive ubiquitin p62/Sqstm1,...
The adaptor protein 1 (AP1) complex is a heterotetramer that participates in cargo sorting into clathrin-coated vesicles at the trans-Golgi network (TGN) and endosomes. gamma subunit of AP1 possesses C-terminal "ear" domain recruits cohort accessory proteins through recognition shared canonical motif, PsiG[PDE][PsiLM] (where Psi an aromatic residue). physiological relevance these ear-motif interactions, however, remains to be demonstrated. Here we report cyclin G-associated kinase (GAK) has...
In addition to the caspase family of proteinases, cathepsin D, a lysosomal aspartic proteinase, has been suggested act as proapoptotic mediator in mammalian cells. To further understand roles cathepsins B and D apoptosis cells, we examined precise alteration processes ultrastructures immunoreactivity for these enzymes PC12 cells cultured under serum deprivation.Laser scanning microscopy showed be finely distributed cytoplasm at onset culture deprivation. At 3h after culture, slightly...
BACE (β‐site amyloid precursor protein cleaving enzyme, β‐secretase) is a type‐I membrane which functions as an aspartic protease in the production of β‐amyloid peptide, causative agent Alzheimer's disease. Its cytoplasmic tail has characteristic acidic‐cluster dileucine motif recognized by VHS domain adaptor proteins, GGAs (Golgi‐localizing, γ‐adaptin ear homology domain, ARF‐interacting). Here we show that colocalized with trans ‐Golgi network and peripheral structures, phosphorylation...
Cachexia is a multifactorial syndrome characterized by muscle loss that cannot be reversed conventional nutritional support. To uncover the molecular basis underlying onset of cancer cachectic wasting and establish an effective intervention against loss, we used mouse model examined effects aerobic exercise. Aerobic exercise successfully suppressed atrophy activated adiponectin signaling. Next, cellular for using C2C12 myotubes was prepared treating with conditioned medium from culture...
Abstract Epidermal growth factor receptor (EGFR) signaling and its downregulation upon ligand binding have been extensively documented. However, the mechanisms by which cells maintain steady-state EGFR expression remain poorly understood. Here, we report a novel role of Golgi-localized, γ-adaptin ear-containing, ADP ribosylation factor-binding protein 2 (GGA2) in control turnover. Whereas GGA1- or GGA3-depletion increased expression, GGA2-depletion RNAi greatly reduced EGFR, reflecting...
Targeting of proteins to their final destination is a prerequisite for living cells maintain homeostasis. Clathrin functions as coat that forms transport carriers called clathrin-coated vesicles (CCVs) at the plasma membrane and post-Golgi compartments. In this study, we established an experimental system using Schneider S2 derived from fruit fly, Drosophila melanogaster, model study physiological roles clathrin adaptors, dissect processes CCV formation. We found adaptor GGA (dGGA), homolog...
Abstract The morphology of the Golgi complex is influenced by cellular context, which strictly correlates with nuclear functions; however, mechanism underlying this association remains elusive. inner membrane SUN proteins, SUN1 and SUN2, have diverse functions together outer nesprin comprise LINC complex. We found that depletion leads to dispersion maintenance ministacks retained function for vesicle transport through In addition, SUN2 associates microtubule plus-end-directed motor KIF20A,...
Gold nanoparticles (AuNPs) can be used with megavolt (MV) X-rays to exert radiosensitization effects, as demonstrated in cell survival assays and mouse experiments. However, the detailed mechanisms are not clear; besides physical dose enhancement, several chemical biological processes have been proposed. Reducing AuNP concentration while achieving sufficient enhancement is necessary for clinical application of AuNPs. Here, we positively charged (+) AuNPs determine effects combined MV on DNA...
Clathrin adaptor protein complex-1 (AP-1) and its accessory proteins play a role in the sorting of integral membrane at trans-Golgi network endosomes. Their physiological functions complex organisms, however, are not fully understood. In this study, we found that CG8538p, an uncharacterized Drosophila protein, shares significant structural functional characteristics with Aftiphilin, mammalian AP-1 protein. The Aftiphilin was shown to interact directly ear domain γ-adaptin AP-1, but GAE GGA....
A live assay tool has been established to uncover the precise molecular mechanisms underlying complex cell fusion events in myoblasts. The novel cell-based assay, HiMy (HiBiT-based myoblast fusion), utilizes a recently developed split-luciferase technology. successfully detected differentiating C2C12 cultures. This allowed us measure mixing of cytoplasm, which occurred several hours after initiation differentiation. Unlike what was reported earlier, few Thus, this is sensitive enough monitor...
To understand the precise distribution of tripeptidyl peptidase I (TPP-I), a defect which has been shown to induce late infantile neuronal ceroid lipofuscinosis, various tissues from rats and mice were analyzed using biochemical immunohistochemical techniques. Western blot analyses showed that protein band immunoreactive anti-TPP-I appeared in tissue extracts both animals at molecular weight approximately 47 kD. Protein levels TPP-I differed among tissues; they high rat brain, liver,...
The Golgi-localized, Gamma-ear-containing, Arf-binding (GGA) proteins are monomeric clathrin adaptors that mediate the sorting of transmembrane cargo at trans-Golgi network and endosomes. Here we report one these proteins, GGA3, becomes transiently phosphorylated upon activation epidermal growth factor (EGF) receptor. This phosphorylation takes place on a previously unrecognized site in "hinge" segment protein, S368, is strictly dependent constitutive another site, S372. EGF-induced S368...
During myogenesis, myogenic stem cells undergo several sequential events, including cell division, migration, and cell-cell fusion, leading to the formation of multinuclear myotubes, which are precursors myofibers. To understand molecular mechanisms underlying these complex processes, an RNA interference-based gene depletion approach was used. Golgi associated, gamma adaptin ear containing, ARF binding protein 1 (GGA1), a Golgi-resident monomeric clathrin adaptor, found be required for...