start-ver=1.4
cd-journal=joma
no-vol=10
cd-vols=
no-issue=1
article-no=
start-page=4798
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2020
dt-pub=20200316
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Development of an experimental method of systematically estimating protein expression limits in HEK293 cells
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Protein overexpression sometimes causes cellular defects, although the underlying mechanism is still unknown. A protein's expression limit, which triggers cellular defects, is a useful indication of the underlying mechanism. In this study, we developed an experimental method of estimating the expression limits of target proteins in the human embryonic kidney cell line HEK293 by measuring the proteins' expression levels in cells that survived after the high-copy introduction of plasmid DNA by which the proteins were expressed under a strong cytomegalovirus promoter. The expression limits of nonfluorescent target proteins were indirectly estimated by measuring the levels of green fluorescent protein (GFP) connected to the target proteins with the self-cleaving sequence P2A. The expression limit of a model GFP was similar to 5.0% of the total protein, and sustained GFP overexpression caused cell death. The expression limits of GFPs with mitochondria-targeting signals and endoplasmic reticulum localization signals were 1.6% and 0.38%, respectively. The expression limits of four proteins involved in vesicular trafficking were far lower compared to a red fluorescent protein. The protein expression limit estimation method developed will be valuable for defining toxic proteins and consequences of protein overexpression.
en-copyright=
kn-copyright=
en-aut-name=MoriYoshihiro
en-aut-sei=Mori
en-aut-mei=Yoshihiro
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=YoshidaYuki
en-aut-sei=Yoshida
en-aut-mei=Yuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=MoriyaHisao
en-aut-sei=Moriya
en-aut-mei=Hisao
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
affil-num=1
en-affil=Graduate School of Natural Science and Technology, Okayama University
kn-affil=
affil-num=2
en-affil=Sony Computer Science Laboratories
kn-affil=
affil-num=3
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=4
en-affil=Research Core for Interdisciplinary Sciences, Okayama University
kn-affil=
en-keyword=Biological techniques
kn-keyword=Biological techniques
en-keyword=Cell biology
kn-keyword=Cell biology
en-keyword=Gene expression analysis
kn-keyword=Gene expression analysis
en-keyword=Molecular biology
kn-keyword=Molecular biology
en-keyword=Protein translocation
kn-keyword=Protein translocation
en-keyword=Protein transport
kn-keyword=Protein transport
END
start-ver=1.4
cd-journal=joma
no-vol=12
cd-vols=
no-issue=5
article-no=
start-page=
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2020
dt-pub=20200516
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Knockout of MMP3 Weakens Solid Tumor Organoids and Cancer Extracellular Vesicles
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=The tumor organoid (tumoroid) model in three-dimensional (3D) culture systems has been developed to reflect more closely the in vivo tumors than 2D-cultured tumor cells. Notably, extracellular vesicles (EVs) are efficiently collectible from the culture supernatant of gel-free tumoroids. Matrix metalloproteinase (MMP) 3 is a multi-functional factor playing crucial roles in tumor progression. However, roles of MMP3 within tumor growth and EVs have not unveiled. Here, we investigated the protumorigenic roles of MMP3 on integrities of tumoroids and EVs. We generated MMP3-knockout (KO) cells using the CRISPR/Cas9 system from rapidly metastatic LuM1 tumor cells. Moreover, we established fluorescent cell lines with palmitoylation signal-fused fluorescent proteins (tdTomato and enhanced GFP). Then we confirmed the exchange of EVs between cellular populations and tumoroids. LuM1-tumoroids released large EVs (200-1000 nm) and small EVs (50-200 nm) while the knockout of MMP3 resulted in the additional release of broken EVs from tumoroids. The loss of MMP3 led to a significant reduction in tumoroid size and the development of the necrotic area within tumoroids. MMP3 and CD9 (a category-1 EV marker tetraspanin protein) were significantly down-regulated in MMP3-KO cells and their EV fraction. Moreover, CD63, another member of the tetraspanin family, was significantly reduced only in the EVs fractions of the MMP3-KO cells compared to their counterpart. These weakened phenotypes of MMP3-KO were markedly rescued by the addition of MMP3-rich EVs or conditioned medium (CM) collected from LuM1-tumoroids, which caused a dramatic rise in the expression of MMP3, CD9, and Ki-67 (a marker of proliferating cells) in the MMP3-null/CD9-low tumoroids. Notably, MMP3 enriched in tumoroids-derived EVs and CM deeply penetrated recipient MMP3-KO tumoroids, resulting in a remarkable enlargement of solid tumoroids, while MMP3-null EVs did not. These data demonstrate that EVs can mediate molecular transfer of MMP3, resulting in increasing the proliferation and tumorigenesis, indicating crucial roles of MMP3 in tumor progression.
en-copyright=
kn-copyright=
en-aut-name=TahaEman A.
en-aut-sei=Taha
en-aut-mei=Eman A.
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=SogawaChiharu
en-aut-sei=Sogawa
en-aut-mei=Chiharu
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=OkushaYuka
en-aut-sei=Okusha
en-aut-mei=Yuka
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=KawaiHotaka
en-aut-sei=Kawai
en-aut-mei=Hotaka
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=OoMay Wathone
en-aut-sei=Oo
en-aut-mei=May Wathone
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=ElseoudiAbdellatif
en-aut-sei=Elseoudi
en-aut-mei=Abdellatif
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=LuYanyin
en-aut-sei=Lu
en-aut-mei=Yanyin
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=NagatsukaHitoshi
en-aut-sei=Nagatsuka
en-aut-mei=Hitoshi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
en-aut-name=KubotaSatoshi
en-aut-sei=Kubota
en-aut-mei=Satoshi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=9
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=10
ORCID=
en-aut-name=OkamotoKuniaki
en-aut-sei=Okamoto
en-aut-mei=Kuniaki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=11
ORCID=
en-aut-name=EguchiTakanori
en-aut-sei=Eguchi
en-aut-mei=Takanori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=12
ORCID=
affil-num=1
en-affil=Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=2
en-affil=Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=3
en-affil=Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=4
en-affil=Department of Oral Pathology and Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=5
en-affil=Department of Oral Pathology and Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=6
en-affil=Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=7
en-affil=Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=8
en-affil=Department of Oral Pathology and Medicine, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=9
en-affil=Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=10
en-affil=Department of Medical Bioengineering, Okayama University Graduate School of Natural Science and Technology
kn-affil=
affil-num=11
en-affil=Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
affil-num=12
en-affil=Department of Dental Pharmacology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
kn-affil=
en-keyword=matrix metalloproteinase 3 (MMP3)
kn-keyword=matrix metalloproteinase 3 (MMP3)
en-keyword=extracellular vesicles (EVs)
kn-keyword=extracellular vesicles (EVs)
en-keyword=tumoroid
kn-keyword=tumoroid
en-keyword=tumor organoid
kn-keyword=tumor organoid
en-keyword=tumorigenesis
kn-keyword=tumorigenesis
en-keyword=three-dimensional (3D) culture system
kn-keyword=three-dimensional (3D) culture system
END
start-ver=1.4
cd-journal=joma
no-vol=12
cd-vols=
no-issue=1
article-no=
start-page=82
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2019
dt-pub=20191229
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Hematopoietic Cells Derived from Cancer Stem Cells Generated from Mouse Induced Pluripotent Stem Cells
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Cancer stem cells (CSCs) represent the subpopulation of cancer cells with the ability to differentiate into other cell phenotypes and initiated tumorigenesis. Previously, we reported generating CSCs from mouse induced pluripotent stem cells (miPSCs). Here, we investigated the ability of the CSCs to differentiate into hematopoietic cells. First, the primary cells were isolated from malignant tumors that were formed by the CSCs. Non-adherent cells (NACs) that arose from adherent cells were collected and their viability, as well as the morphology and expression of hematopoietic cell markers, were analyzed. Moreover, NACs were injected into the tail vein of busulfan conditioned Balb/c nude mice. Finally, CSCs were induced to differentiate to macrophages while using IL3 and SCF. The round nucleated NACs were found to be viable, positive for hematopoietic lineage markers and CD34, and expressed hematopoietic markers, just like homing to the bone marrow. When NACs were injected into mice, Wright-Giemsa staining showed that the number of white blood cells got higher than those in the control mice after four weeks. CSCs also showed the ability to differentiate toward macrophages. CSCs were demonstrated to have the potential to provide progenies with hematopoietic markers, morphology, and homing ability to the bone marrow, which could give new insight into the tumor microenvironment according to the plasticity of CSCs.
en-copyright=
kn-copyright=
en-aut-name=HassanGhmkin
en-aut-sei=Hassan
en-aut-mei=Ghmkin
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=AfifySaid M.
en-aut-sei=Afify
en-aut-mei=Said M.
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=NairNeha
en-aut-sei=Nair
en-aut-mei=Neha
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=KumonKazuki
en-aut-sei=Kumon
en-aut-mei=Kazuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=OsmanAmira
en-aut-sei=Osman
en-aut-mei=Amira
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=DuJuan
en-aut-sei=Du
en-aut-mei=Juan
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=MansourHager
en-aut-sei=Mansour
en-aut-mei=Hager
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=Abu QuoraHagar A.
en-aut-sei=Abu Quora
en-aut-mei=Hagar A.
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
en-aut-name=NawaraHend M.
en-aut-sei=Nawara
en-aut-mei=Hend M.
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=9
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=10
ORCID=
en-aut-name=ZahraMaram H.
en-aut-sei=Zahra
en-aut-mei=Maram H.
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=11
ORCID=
en-aut-name=OkadaNobuhiro
en-aut-sei=Okada
en-aut-mei=Nobuhiro
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=12
ORCID=
en-aut-name=SenoAkimasa
en-aut-sei=Seno
en-aut-mei=Akimasa
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=13
ORCID=
en-aut-name=SenoMasaharu
en-aut-sei=Seno
en-aut-mei=Masaharu
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=14
ORCID=
affil-num=1
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=2
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=3
en-affil=Graduate School of Natural Science and Technology, Okayama University
kn-affil=
affil-num=4
en-affil=Graduate School of Natural Science and Technology, Okayama University
kn-affil=
affil-num=5
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=6
en-affil=Graduate School of Natural Science and Technology, Okayama University
kn-affil=
affil-num=7
en-affil=
kn-affil=Graduate School of Natural Science and Technology, Okayama University
affil-num=8
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=9
en-affil=Graduate School of Natural Science and Technology, Okayama University
kn-affil=
affil-num=10
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=11
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=12
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=13
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=14
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
en-keyword=Induced pluripotent stem cells
kn-keyword=Induced pluripotent stem cells
en-keyword=Cancer stem cells differentiation
kn-keyword=Cancer stem cells differentiation
en-keyword=tumor microenvironment
kn-keyword=tumor microenvironment
en-keyword=hematopoietic cells
kn-keyword=hematopoietic cells
END
start-ver=1.4
cd-journal=joma
no-vol=9
cd-vols=
no-issue=
article-no=
start-page=8866
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2019
dt-pub=2019620
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Yeast screening system reveals the inhibitory mechanism of cancer cell proliferation by benzyl isothiocyanate through down-regulation of Mis12
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Benzyl isothiocyanate (BITC) is a naturally-occurring isothiocyanate derived from cruciferous vegetables. BITC has been reported to inhibit the proliferation of various cancer cells, which is believed to be important for the inhibition of tumorigenesis. However, the detailed mechanisms of action remain unclear. In this study, we employed a budding yeast Saccharomyces cerevisiae as a model organism for screening. Twelve genes including MTW1 were identified as the overexpression suppressors for the antiproliferative effect of BITC using the genome-wide multi-copy plasmid collection for S. cerevisiae. Overexpression of the kinetochore protein Mtw1 counteracts the antiproliferative effect of BITC in yeast. The inhibitory effect of BITC on the proliferation of human colon cancer HCT-116 cells was consistently suppressed by the overexpression of Mis12, a human orthologue of Mtw1, and enhanced by the knockdown of Mis12. We also found that BITC increased the phosphorylated and ubiquitinated Mis12 level with consequent reduction of Mis12, suggesting that BITC degrades Mis12 through an ubiquitin-proteasome system. Furthermore, cell cycle analysis showed that the change in the Mis12 level affected the cell cycle distribution and the sensitivity to the BITC-induced apoptosis. These results provide evidence that BITC suppresses cell proliferation through the post-transcriptional regulation of the kinetochore protein Mis12.
en-copyright=
kn-copyright=
en-aut-name=Abe-KanohNaomi
en-aut-sei=Abe-Kanoh
en-aut-mei=Naomi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=KunisueNarumi
en-aut-sei=Kunisue
en-aut-mei=Narumi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=MyojinTakumi
en-aut-sei=Myojin
en-aut-mei=Takumi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=ChinoAyako
en-aut-sei=Chino
en-aut-mei=Ayako
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=MunemasaShintaro
en-aut-sei=Munemasa
en-aut-mei=Shintaro
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=MurataYoshiyuki
en-aut-sei=Murata
en-aut-mei=Yoshiyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=MoriyaHisao
en-aut-sei=Moriya
en-aut-mei=Hisao
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
en-aut-name=NakamuraYoshimasa
en-aut-sei=Nakamura
en-aut-mei=Yoshimasa
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=9
ORCID=
affil-num=1
en-affil=Graduate School of Environmental and Life Science, Okayama University
kn-affil=
affil-num=2
en-affil=Graduate School of Environmental and Life Science, Okayama University
kn-affil=
affil-num=3
en-affil=Graduate School of Environmental and Life Science, Okayama University
kn-affil=
affil-num=4
en-affil=Research Core for Interdisciplinary Sciences, Okayama University
kn-affil=
affil-num=5
en-affil=Graduate School of Environmental and Life Science, Okayama University
kn-affil=
affil-num=6
en-affil=Graduate School of Environmental and Life Science, Okayama University
kn-affil=
affil-num=7
en-affil= Graduate School of Natural Science and Technology, Okayama University
kn-affil=
affil-num=8
en-affil=Research Core for Interdisciplinary Sciences, Okayama University
kn-affil=
affil-num=9
en-affil=Graduate School of Environmental and Life Science, Okayama University, Okayama
kn-affil=
END
start-ver=1.4
cd-journal=joma
no-vol=7
cd-vols=
no-issue=
article-no=
start-page=160
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2019
dt-pub=20190827
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=The Golgin Protein Giantin Regulates Interconnections Between Golgi Stacks
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract= Golgins are a family of Golgi-localized long coiled-coil proteins. The major golgin function is thought to be the tethering of vesicles, membranes, and cytoskeletal elements to the Golgi. We previously showed that knockdown of one of the longest golgins, Giantin, altered the glycosylation patterns of cell surfaces and the kinetics of cargo transport, suggesting that Giantin maintains correct glycosylation through slowing down transport within the Golgi. Giantin knockdown also altered the sizes and numbers of mini Golgi stacks generated by microtubule de-polymerization, suggesting that it maintains the independence of individual Golgi stacks. Therefore, it is presumed that Golgi stacks lose their independence following Giantin knockdown, allowing easier and possibly increased transport among stacks and abnormal glycosylation. To gain structural insights into the independence of Golgi stacks, we herein performed electron tomography and 3D modeling of Golgi stacks in Giantin knockdown cells. Compared with control cells, Giantin-knockdown cells had fewer and smaller fenestrae within each cisterna. This was supported by data showing that the diffusion rate of Golgi membrane proteins is faster in Giantin-knockdown Golgi, indicating that Giantin knockdown structurally and functionally increases connectivity among Golgi cisternae and stacks. This increased connectivity suggests that contrary to the cis-golgin tether model, Giantin instead inhibits the tether and fusion of nearby Golgi cisternae and stacks, resulting in transport difficulties between stacks that may enable the correct glycosylation of proteins and lipids passing through the Golgi.
en-copyright=
kn-copyright=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=Hayashi-NishinoMitsuko
en-aut-sei=Hayashi-Nishino
en-aut-mei=Mitsuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=ShakunoTakuto
en-aut-sei=Shakuno
en-aut-mei=Takuto
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=MasudaJunko
en-aut-sei=Masuda
en-aut-mei=Junko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=KoreishiMayuko
en-aut-sei=Koreishi
en-aut-mei=Mayuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=MurakamiRuna
en-aut-sei=Murakami
en-aut-mei=Runa
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=NakamuraYoshimasa
en-aut-sei=Nakamura
en-aut-mei=Yoshimasa
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=NakamuraToshiyuki
en-aut-sei=Nakamura
en-aut-mei=Toshiyuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
en-aut-name=Abe-KanohNaomi
en-aut-sei=Abe-Kanoh
en-aut-mei=Naomi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=9
ORCID=
en-aut-name=HonjoYasuko
en-aut-sei=Honjo
en-aut-mei=Yasuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=10
ORCID=
en-aut-name=MalsamJoerg
en-aut-sei=Malsam
en-aut-mei=Joerg
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=11
ORCID=
en-aut-name=YuSidney
en-aut-sei=Yu
en-aut-mei=Sidney
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=12
ORCID=
en-aut-name=ishinoKunihiko
en-aut-sei=ishino
en-aut-mei=Kunihiko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=13
ORCID=
affil-num=1
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=2
en-affil=Institute of Scientific and Industrial Research, Osaka University
kn-affil=
affil-num=3
en-affil=Graduate School of Natural Science and Technology, Okayama University
kn-affil=
affil-num=4
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=5
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=6
en-affil=Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
kn-affil=
affil-num=7
en-affil=raduate School of Environmental and Life Science, Okayama University
kn-affil=
affil-num=8
en-affil=raduate School of Environmental and Life Science, Okayama University
kn-affil=
affil-num=9
en-affil=raduate School of Environmental and Life Science, Okayama University
kn-affil=
affil-num=10
en-affil=esearch Institute for Radiation Biology and Medicine, Hiroshima University
kn-affil=
affil-num=11
en-affil=Center for Biochemistry (BZH), Heidelberg University
kn-affil=
affil-num=12
en-affil=School of Biomedical Sciences, The Chinese University of Hong Kong
kn-affil=
affil-num=13
en-affil=Institute of Scientific and Industrial Research, Osaka University
kn-affil=
en-keyword=Golgi
kn-keyword=Golgi
en-keyword=golgins
kn-keyword=golgins
en-keyword=glycosylation
kn-keyword=glycosylation
en-keyword=endoplasmic reticulum
kn-keyword=endoplasmic reticulum
en-keyword=electron tomography
kn-keyword=electron tomography
END
start-ver=1.4
cd-journal=joma
no-vol=25
cd-vols=
no-issue=146
article-no=
start-page=241
end-page=244
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2013
dt-pub=201311
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=ゴルジ体内の輸送：糖タンパク質の移動に関する研究
kn-title=Transport within the Golgi: for the Study of Glycoprotein Movement
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=
en-copyright=
kn-copyright=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=佐藤あやの
kn-aut-sei=佐藤
kn-aut-mei=あやの
aut-affil-num=1
ORCID=
en-aut-name=IwakiriYasuko
en-aut-sei=Iwakiri
en-aut-mei=Yasuko
kn-aut-name=岩切泰子
kn-aut-sei=岩切
kn-aut-mei=泰子
aut-affil-num=2
ORCID=
affil-num=1
en-affil=
kn-affil=岡山大学大学院自然科学研究科
affil-num=2
en-affil=
kn-affil=エール大学医学部内科消化器科
en-keyword=Golgi
kn-keyword=Golgi
en-keyword=glycosylation
kn-keyword=glycosylation
en-keyword=intracellular trafficking
kn-keyword=intracellular trafficking
END
start-ver=1.4
cd-journal=joma
no-vol=182
cd-vols=
no-issue=3
article-no=
start-page=786
end-page=795
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2013
dt-pub=201303
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Absence of Nogo-B (Reticulon 4B) Facilitates Hepatic Stellate Cell Apoptosis and Diminishes Hepatic Fibrosis in Mice
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Nogo-B (reticulon 4B) accentuates hepatic fibrosis and cirrhosis, but the mechanism remains unclear. The aim of this study was to identify the role of Nogo-B in hepatic stellate cell (HSC) apoptosis in cirrhotic livers. Cirrhosis was generated by carbon tetrachloride inhalation in wild-type (WT) and Nogo-A/B knockout (Nogo-B KO) mice. HSCs were isolated from WT and Nogo-B KO mice and cultured for activation and transformation to myofibroblasts (MF-HSCs). Human hepatic stellate cells (LX2 cells) were used to assess apoptotic responses of activated HSCs after silencing or overexpressing Nogo-B. Livers from cirrhotic Nogo-B KO mice showed significantly reduced fibrosis (P < 0.05) compared with WT mice. Apoptotic cells were more prominent in fibrotic areas of cirrhotic Nogo-B KO livers. Nogo-B KO MF-HSCs showed significantly increased Levels of apoptotic markers, cleaved poly (ADP-ribose) polymerase, and caspase-3 and -8 (P < 0.05) compared with WT MF-HSCs in response to staurosporine. Treatment with tunicamycin, an endoplasmic reticulum stress inducer, increased cleaved caspase-3 and -8 levels in Nogo-B KO MF-HSCs compared with WT MF-HSCs (P < 0.01). In LX2 cells, Nogo-B knockdown enhanced apoptosis in response to staurosporine, whereas Nogo-B overexpression inhibited apoptosis. The absence of Nogo-B enhances apoptosis of HSCs in experimental cirrhosis. Selective blockade of Nogo-B in HSCs may represent a potential therapeutic strategy to mitigate liver fibrosis. (Am J Pathol 2013, 182: 786-795; http://dx.doLorg/10.1016Aajpath.2012.11.032)
en-copyright=
kn-copyright=
en-aut-name=TashiroKeitaro
en-aut-sei=Tashiro
en-aut-mei=Keitaro
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=UtsumiTeruo
en-aut-sei=Utsumi
en-aut-mei=Teruo
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=ChungChuhan
en-aut-sei=Chung
en-aut-mei=Chuhan
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=IwakiriYasuko
en-aut-sei=Iwakiri
en-aut-mei=Yasuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
affil-num=1
en-affil=
kn-affil=Yale Univ, Sch Med, Sect Digest Dis, Dept Internal Med
affil-num=2
en-affil=
kn-affil=Okayama Univ, Grad Sch Nat Sci
affil-num=3
en-affil=
kn-affil=Yale Univ, Sch Med, Sect Digest Dis, Dept Internal Med
affil-num=4
en-affil=
kn-affil=Yale Univ, Sch Med, Sect Digest Dis, Dept Internal Med
affil-num=5
en-affil=
kn-affil=Yale Univ, Sch Med, Sect Digest Dis, Dept Internal Med
END
start-ver=1.4
cd-journal=joma
no-vol=8
cd-vols=
no-issue=3
article-no=
start-page=
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2013
dt-pub=20130521
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=The Golgin Tether Giantin Regulates the Secretory Pathway by Controlling Stack Organization within Golgi Apparatus
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Golgins are coiled-coil proteins that play a key role in the regulation of Golgi architecture and function. Giantin, the largest golgin in mammals, forms a complex with p115, rab1, GM130, and soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), thereby facilitating vesicle tethering and fusion processes around the Golgi apparatus. Treatment with the microtubule destabilizing drug nocodazole transforms the Golgi ribbon into individual Golgi stacks. Here we show that siRNA-mediated depletion of giantin resulted in more dispersed Golgi stacks after nocodazole treatment than by control treatment, without changing the average cisternal length. Furthermore, depletion of giantin caused an increase in cargo transport that was associated with altered cell surface protein glycosylation. Drosophila S2 cells are known to have dispersed Golgi stacks and no giantin homolog. The exogenous expression of mammalian giantin cDNA in S2 cells resulted in clustered Golgi stacks, similar to the Golgi ribbon in mammalian cells. These results suggest that the spatial organization of the Golgi ribbon is mediated by giantin, which also plays a role in cargo transport and sugar modifications.
en-copyright=
kn-copyright=
en-aut-name=KoreishiMayuko
en-aut-sei=Koreishi
en-aut-mei=Mayuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=GniadekThomas J.
en-aut-sei=Gniadek
en-aut-mei=Thomas J.
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=YuSidney
en-aut-sei=Yu
en-aut-mei=Sidney
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=MasudaJunko
en-aut-sei=Masuda
en-aut-mei=Junko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=HonjoYasuko
en-aut-sei=Honjo
en-aut-mei=Yasuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
affil-num=1
en-affil=
kn-affil=Okayama Univ, Grad Sch Nat Sci & Technol
affil-num=2
en-affil=
kn-affil=Johns Hopkins Univ, Sch Med, Dept Pathol
affil-num=3
en-affil=
kn-affil=Chinese Univ Hong Kong, Sch Biomed Sci, Shatin
affil-num=4
en-affil=
kn-affil=NIAID, Mucosal Immun Sect, Lab Host Def
affil-num=5
en-affil=
kn-affil=Okayama Univ, RCIS
affil-num=6
en-affil=
kn-affil=Okayama Univ, Grad Sch Nat Sci & Technol
END
start-ver=1.4
cd-journal=joma
no-vol=8
cd-vols=
no-issue=1
article-no=
start-page=
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2013
dt-pub=20130118
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=CK2 Phosphorylates Sec31 and Regulates ER-To-Golgi Trafficking
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Protein export from the endoplasmic reticulum (ER) is an initial and rate-limiting step of molecular trafficking and secretion. This is mediated by coat protein II (COPII)-coated vesicles, whose formation requires small GTPase Sar1 and 6 Sec proteins including Sec23 and Sec31. Sec31 is a component of the outer layer of COPII coat and has been identified as a phosphoprotein. The initiation and promotion of COPII vesicle formation is regulated by Sar1; however, the mechanism regulating the completion of COPII vesicle formation followed by vesicle release is largely unknown. Hypothesizing that the Sec31 phosphorylation may be such a mechanism, we identified phosphorylation sites in the middle linker region of Sec31. Sec31 phosphorylation appeared to decrease its association with ER membranes and Sec23. Non-phosphorylatable mutant of Sec31 stayed longer at ER exit sites and bound more strongly to Sec23. We also found that CK2 is one of the kinases responsible for Sec31 phosphorylation because CK2 knockdown decreased Sec31 phosphorylation, whereas CK2 overexpression increased Sec31 phosphorylation. Furthermore, CK2 knockdown increased affinity of Sec31 for Sec23 and inhibited ER-to-Golgi trafficking. These results suggest that Sec31 phosphorylation by CK2 controls the duration of COPII vesicle formation, which regulates ER-to-Golgi trafficking.
en-copyright=
kn-copyright=
en-aut-name=KoreishiMayuko
en-aut-sei=Koreishi
en-aut-mei=Mayuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=YuSidney
en-aut-sei=Yu
en-aut-mei=Sidney
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=OdaMayumi
en-aut-sei=Oda
en-aut-mei=Mayumi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=HonjoYasuko
en-aut-sei=Honjo
en-aut-mei=Yasuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
affil-num=1
en-affil=
kn-affil=Okayama Univ, Grad Sch Nat Sci & Technol
affil-num=2
en-affil=
kn-affil=Chinese Univ Hong Kong, Sch Biomed Sci
affil-num=3
en-affil=
kn-affil=Okayama Univ, Grad Sch Nat Sci & Technol
affil-num=4
en-affil=
kn-affil=Okayama Univ, Res Core Interdisciplinary Sci RCIS
affil-num=5
en-affil=
kn-affil=Okayama Univ, Grad Sch Nat Sci & Technol
END
start-ver=1.4
cd-journal=joma
no-vol=7
cd-vols=
no-issue=4
article-no=
start-page=
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2012
dt-pub=20120412
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=A Model of Cancer Stem Cells Derived from Mouse Induced Pluripotent Stem Cells
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Cancer stem cells (CSCs) are capable of continuous proliferation and self-renewal and are proposed to play significant roles in oncogenesis, tumor growth, metastasis and cancer recurrence. CSCs are considered derived from normal stem cells affected by the tumor microenvironment although the mechanism of development is not clear yet. In 2007, Yamanaka's group succeeded in generating Nanog mouse induced pluripotent stem (miPS) cells, in which green fluorescent protein (GFP) has been inserted into the 5'-untranslated region of the Nanog gene. Usually, iPS cells, just like embryonic stem cells, are considered to be induced into progenitor cells, which differentiate into various normal phenotypes depending on the normal niche. We hypothesized that CSCs could be derived from Nanog miPS cells in the conditioned culture medium of cancer cell lines, which is a mimic of carcinoma microenvironment. As a result, the Nanog miPS cells treated with the conditioned medium of mouse Lewis lung carcinoma acquired characteristics of CSCs, in that they formed spheroids expressing GFP in suspension culture, and had a high tumorigenicity in Balb/c nude mice exhibiting angiogenesis in vivo. In addition, these iPS-derived CSCs had a capacity of self-renewal and expressed the marker genes, Nanog, Rex1, Eras, Esg1 and Cripto, associated with stem cell properties and an undifferentiated state. Thus we concluded that a model of CSCs was originally developed from miPS cells and proposed the conditioned culture medium of cancer cell lines might perform as niche for producing CSCs. The model of CSCs and the procedure of their establishment will help study the genetic alterations and the secreted factors in the tumor microenvironment which convert miPS cells to CSCs. Furthermore, the identification of potentially bona fide markers of CSCs, which will help the development of novel anti-cancer therapies, might be possible though the CSC model.
en-copyright=
kn-copyright=
en-aut-name=ChenLing
en-aut-sei=Chen
en-aut-mei=Ling
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=KasaiTomonari
en-aut-sei=Kasai
en-aut-mei=Tomonari
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=LiYueguang
en-aut-sei=Li
en-aut-mei=Yueguang
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=SugiiYuh
en-aut-sei=Sugii
en-aut-mei=Yuh
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=JinGuoliang
en-aut-sei=Jin
en-aut-mei=Guoliang
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=OkadaMasashi
en-aut-sei=Okada
en-aut-mei=Masashi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=VaidyanathArun
en-aut-sei=Vaidyanath
en-aut-mei=Arun
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=MizutaniAkifumi
en-aut-sei=Mizutani
en-aut-mei=Akifumi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=9
ORCID=
en-aut-name=KudohTakayuki
en-aut-sei=Kudoh
en-aut-mei=Takayuki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=10
ORCID=
en-aut-name=HendrixMary J. C.
en-aut-sei=Hendrix
en-aut-mei=Mary J. C.
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=11
ORCID=
en-aut-name=SalomonDavid S
en-aut-sei=Salomon
en-aut-mei=David S
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=12
ORCID=
en-aut-name=FuLi
en-aut-sei=Fu
en-aut-mei=Li
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=13
ORCID=
en-aut-name=SenoMasaharu
en-aut-sei=Seno
en-aut-mei=Masaharu
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=14
ORCID=
affil-num=1
en-affil=
kn-affil=Okayama Univ
affil-num=2
en-affil=
kn-affil=Okayama Univ
affil-num=3
en-affil=
kn-affil=Tianjin 4th Ctr Hosp
affil-num=4
en-affil=
kn-affil=Okayama Univ
affil-num=5
en-affil=
kn-affil=Okayama Univ
affil-num=6
en-affil=
kn-affil=Okayama Univ
affil-num=7
en-affil=
kn-affil=Okayama Univ
affil-num=8
en-affil=
kn-affil=Okayama Univ
affil-num=9
en-affil=
kn-affil=Okayama Univ
affil-num=10
en-affil=
kn-affil=Okayama Univ
affil-num=11
en-affil=
kn-affil=Northwestern Univ
affil-num=12
en-affil=
kn-affil=NCI
affil-num=13
en-affil=
kn-affil=Tianjin Med Univ
affil-num=14
en-affil=
kn-affil=Okayama Univ
END
start-ver=1.4
cd-journal=joma
no-vol=2011
cd-vols=
no-issue=1
article-no=
start-page=5
end-page=11
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=201-
dt-pub=2010809
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=Detection of in situ cleaved p115 with the cut specific antibodies in rapid protein inactivation system by tobacco etch viral protease cleavage
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Gene perturbation methods are commonly used in the study of gene and protein function. The authors of this paper recently developed a rapid protein inactivation technique utilizing tobacco etch virus (TEV)-derived protease. TEV protease recognizes the ENLYFQG (Glu-Asn-Leu-Tyr-Phe-Gln-Gly) amino acid sequence and specifically cleaves between Q and G. The authors developed antibodies that recognize the cleaved TEV (ENLYFQ) sequence, both in vitro and in vivo, but do not bind to uncleaved TEV (ENLYFQG). Using these antibodies, in situ protein cleavage was successfully detected. These antibodies used in combination with the TEV protease may be a useful complement to other perturbation methods.
en-copyright=
kn-copyright=
en-aut-name=KoreishiMayuko
en-aut-sei=Koreishi
en-aut-mei=Mayuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=HonjoYasuko
en-aut-sei=Honjo
en-aut-mei=Yasuko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
affil-num=1
en-affil=
kn-affil=The Research Core for Interdisciplinary Sciences (RCIS), Okayama University
affil-num=2
en-affil=
kn-affil=The Research Core for Interdisciplinary Sciences (RCIS), Okayama University
affil-num=3
en-affil=
kn-affil=The Research Core for Interdisciplinary Sciences (RCIS), Okayama University
en-keyword=TEV protease
kn-keyword=TEV protease
en-keyword=Golgi
kn-keyword=Golgi
en-keyword=golgins
kn-keyword=golgins
en-keyword=microinjection
kn-keyword=microinjection
en-keyword=recombinant proteins
kn-keyword=recombinant proteins
END
start-ver=1.4
cd-journal=joma
no-vol=7
cd-vols=
no-issue=1
article-no=
start-page=
end-page=
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2012
dt-pub=20120118
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=TRAPPC9 Mediates the Interaction between p150Glued and COPII Vesicles at the Target Membrane
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Background: The transport of endoplasmic reticulum (ER)-derived COPII vesicles toward the ER-Golgi intermediate
compartment (ERGIC) requires cytoplasmic dynein and is dependent on microtubules. p150Glued, a subunit of dynactin, has
been implicated in the transport of COPII vesicles via its interaction with COPII coat components Sec23 and Sec24. However,
whether and how COPII vesicle tether, TRAPP (Transport protein particle), plays a role in the interaction between COPII
vesicles and microtubules is currently unknown.
Principle Findings: We address the functional relationship between COPII tether TRAPP and dynactin. Overexpressed TRAPP
subunits interfered with microtubule architecture by competing p150Glued away from the MTOC. TRAPP subunit TRAPPC9
bound directly to p150Glued via the same carboxyl terminal domain of p150Glued that binds Sec23 and Sec24. TRAPPC9 also
inhibited the interaction between p150Glued and Sec23/Sec24 both in vitro and in vivo, suggesting that TRAPPC9 serves to
uncouple p150Glued from the COPII coat, and to relay the vesicle-dynactin interaction at the target membrane.
Conclusions: These findings provide a new perspective on the function of TRAPP as an adaptor between the ERGIC
membrane and dynactin. By preserving the connection between dynactin and the tethered and/or fused vesicles, TRAPP
allows nascent ERGIC to continue the movement along the microtubules as they mature into the cis-Golgi.
en-copyright=
kn-copyright=
en-aut-name=ZongMin
en-aut-sei=Zong
en-aut-mei=Min
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=YuMei Kuen
en-aut-sei=Yu
en-aut-mei=Mei Kuen
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=SiuKa Yu
en-aut-sei=Siu
en-aut-mei=Ka Yu
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=NgWing Yan
en-aut-sei=Ng
en-aut-mei=Wing Yan
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=ChaｎHsiao Chang
en-aut-sei=Chaｎ
en-aut-mei=Hsiao Chang
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=TannerJulian A.
en-aut-sei=Tanner
en-aut-mei=Julian A.
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=YuSidney
en-aut-sei=Yu
en-aut-mei=Sidney
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
affil-num=1
en-affil=
kn-affil=School of Biomedical Sciences, The Chinese University of Hong Kong
affil-num=2
en-affil=
kn-affil=The Research Core for Interdisciplinary Science, Okayama University
affil-num=3
en-affil=
kn-affil=Epithelial Cell Biology Research Center, The Chinese University of Hong Kong,
affil-num=4
en-affil=
kn-affil=School of Biomedical Sciences, The Chinese University of Hong Kong
affil-num=5
en-affil=
kn-affil=School of Biomedical Sciences, The Chinese University of Hong Kong
affil-num=6
en-affil=
kn-affil=School of Biomedical Sciences, The Chinese University of Hong Kong
affil-num=7
en-affil=
kn-affil=Department of Biochemistry, The University of Hong Kong
affil-num=8
en-affil=
kn-affil=School of Biomedical Sciences, The Chinese University of Hong Kong
END
start-ver=1.4
cd-journal=joma
no-vol=20
cd-vols=
no-issue=19
article-no=
start-page=4205
end-page=4215
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2009
dt-pub=20091001
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=mTrs130 Is a Component of a Mammalian TRAPPII Complex, a Rab1 GEF That Binds to COPI-coated Vesicles
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=The GTPase Rab1 regulates endoplasmic reticulum-Golgi and early Golgi traffic. The guanine nucleotide exchange factor (GEF) or factors that activate Rab1 at these stages of the secretory pathway are currently unknown. Trs130p is a subunit of the yeast TRAPPII (transport protein particle II) complex, a multisubunit tethering complex that is a GEF for the Rab1 homologue Ypt1p. Here, we show that mammalian Trs130 (mTrs130) is a component of an analogous TRAPP complex in mammalian cells, and we describe for the first time the role that this complex plays in membrane traffic. mTRAPPII is enriched on COPI (Coat Protein I)-coated vesicles and buds, but not Golgi cisternae, and it specifically activates Rab1. In addition, we find that mTRAPPII binds to γ1COP, a COPI coat adaptor subunit. The depletion of mTrs130 by short hairpin RNA leads to an increase of vesicles in the vicinity of the Golgi and the accumulation of cargo in an early Golgi compartment. We propose that mTRAPPII is a Rab1 GEF that tethers COPI-coated vesicles to early Golgi membranes.
en-copyright=
kn-copyright=
en-aut-name=YamasakiAkinori
en-aut-sei=Yamasaki
en-aut-mei=Akinori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=MenonShekar
en-aut-sei=Menon
en-aut-mei=Shekar
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=YuSidney
en-aut-sei=Yu
en-aut-mei=Sidney
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=BarrowmanJemima
en-aut-sei=Barrowman
en-aut-mei=Jemima
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=MeerlooTimo
en-aut-sei=Meerloo
en-aut-mei=Timo
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=OorschotViola
en-aut-sei=Oorschot
en-aut-mei=Viola
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=KlumpermanJudith
en-aut-sei=Klumperman
en-aut-mei=Judith
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=SatohAyano
en-aut-sei=Satoh
en-aut-mei=Ayano
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
en-aut-name=Ferro-NovickSusan
en-aut-sei=Ferro-Novick
en-aut-mei=Susan
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=9
ORCID=
affil-num=1
en-affil=
kn-affil=Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine
affil-num=2
en-affil=
kn-affil=Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine
affil-num=3
en-affil=
kn-affil=Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine
affil-num=4
en-affil=
kn-affil=Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine
affil-num=5
en-affil=
kn-affil=Department of Cellular and Molecular Medicine, University of California at San Diego
affil-num=6
en-affil=
kn-affil=Cell Microscopy Center, Department of Cell Biology, University Medical Center Utrecht
affil-num=7
en-affil=
kn-affil=Cell Microscopy Center, Department of Cell Biology, University Medical Center Utrecht
affil-num=8
en-affil=
kn-affil=Department of Cell Biology, Yale University School of Medicine
affil-num=9
en-affil=
kn-affil=Department of Cell Biology, Howard Hughes Medical Institute, Yale University School of Medicine
END