2022/11/26 更新

写真a

ヨシダ サトシ
吉田 知史
Scopus 論文情報  
論文数: 0  Citation: 0  h-index: 10

Citation Countは当該年に発表した論文の被引用数

所属
国際学術院 国際教養学部
職名
教授

他学部・他研究科等兼任情報

  • 理工学術院   大学院先進理工学研究科

学内研究所・附属機関兼任歴

  • 2021年
    -
    2022年

    リサーチイノベ オープンイノベーション推進セクション   兼任センター員

学歴

  • 1998年
    -
    2003年

    東京大学   大学院理学系研究科  

  • 1994年
    -
    1998年

    東京大学   理学部  

学位

  • 東京大学   博士(理学)

経歴

  • 2020年
    -
    継続中

    早稲田大学   国際教養学部   教授

  • 2018年
    -
    2020年

    早稲田大学   国際教養学部   准教授

  • 2014年
    -
    2018年

    群馬大学   未来先端研究機構   准教授

  • 2009年
    -
    2014年

    ブランダイス大学   生物学部   アシスタントプロフェッサー

  • 2003年
    -
    2009年

    ハーバード大学医学部   研究員

所属学協会

  •  
     
     

    日本分子生物学会

  •  
     
     

    酵母遺伝学フォーラム

 

研究分野

  • 遺伝学   酵母遺伝学

  • 細胞生物学   細胞周期 代謝 老化

研究キーワード

  • 酵母遺伝学

  • 細胞生物学

論文

  • High and stable ATP levels prevent aberrant intracellular protein aggregation in yeast.

    Masak Takaine, Hiromi Imamura, Satoshi Yoshida

    eLife   11  2022年04月  [国際誌]

    担当区分:最終著者, 責任著者

     概要を見る

    Adenosine triphosphate (ATP) at millimolar levels has recently been implicated in the solubilization of cellular proteins. However, the significance of this high ATP level under physiological conditions and the mechanisms that maintain ATP remain unclear. We herein demonstrated that AMP-activated protein kinase (AMPK) and adenylate kinase (ADK) cooperated to maintain cellular ATP levels regardless of glucose levels. Single-cell imaging of ATP-reduced yeast mutants revealed that ATP levels in these mutants underwent stochastic and transient depletion, which promoted the cytotoxic aggregation of endogenous proteins and pathogenic proteins, such as huntingtin and α-synuclein. Moreover, pharmacological elevations in ATP levels in an ATP-reduced mutant prevented the accumulation of α-synuclein aggregates and its cytotoxicity. The present study demonstrates that cellular ATP homeostasis ensures proteostasis and revealed that suppressing the high volatility of cellular ATP levels prevented cytotoxic protein aggregation, implying that AMPK and ADK are important factors that prevent proteinopathies, such as neurodegenerative diseases.

    DOI PubMed

    Scopus

    2
    被引用数
    (Scopus)
  • High-throughput platform for yeast morphological profiling predicts the targets of bioactive compounds.

    Shinsuke Ohnuki, Itsuki Ogawa, Kaori Itto-Nakama, Fachuang Lu, Ashish Ranjan, Mehdi Kabbage, Abraham Abera Gebre, Masao Yamashita, Sheena C Li, Yoko Yashiroda, Satoshi Yoshida, Takeo Usui, Jeff S Piotrowski, Brenda J Andrews, Charles Boone, Grant W Brown, John Ralph, Yoshikazu Ohya

    NPJ systems biology and applications   8 ( 1 ) 3 - 3  2022年01月  [国際誌]

     概要を見る

    Morphological profiling is an omics-based approach for predicting intracellular targets of chemical compounds in which the dose-dependent morphological changes induced by the compound are systematically compared to the morphological changes in gene-deleted cells. In this study, we developed a reliable high-throughput (HT) platform for yeast morphological profiling using drug-hypersensitive strains to minimize compound use, HT microscopy to speed up data generation and analysis, and a generalized linear model to predict targets with high reliability. We first conducted a proof-of-concept study using six compounds with known targets: bortezomib, hydroxyurea, methyl methanesulfonate, benomyl, tunicamycin, and echinocandin B. Then we applied our platform to predict the mechanism of action of a novel diferulate-derived compound, poacidiene. Morphological profiling of poacidiene implied that it affects the DNA damage response, which genetic analysis confirmed. Furthermore, we found that poacidiene inhibits the growth of phytopathogenic fungi, implying applications as an effective antifungal agent. Thus, our platform is a new whole-cell target prediction tool for drug discovery.

    DOI PubMed

    Scopus

  • Defining Functions of Mannoproteins in Saccharomyces cerevisiae by High-Dimensional Morphological Phenotyping.

    Farzan Ghanegolmohammadi, Hiroki Okada, Yaxuan Liu, Kaori Itto-Nakama, Shinsuke Ohnuki, Anna Savchenko, Erfei Bi, Satoshi Yoshida, Yoshikazu Ohya

    Journal of fungi (Basel, Switzerland)   7 ( 9 )  2021年09月  [国際誌]

     概要を見る

    Mannoproteins are non-filamentous glycoproteins localized to the outermost layer of the yeast cell wall. The physiological roles of these structural components have not been completely elucidated due to the limited availability of appropriate tools. As the perturbation of mannoproteins may affect cell morphology, we investigated mannoprotein mutants in Saccharomyces cerevisiae via high-dimensional morphological phenotyping. The mannoprotein mutants were morphologically classified into seven groups using clustering analysis with Gaussian mixture modeling. The pleiotropic phenotypes of cluster I mutant cells (ccw12Δ) indicated that CCW12 plays major roles in cell wall organization. Cluster II (ccw14Δ, flo11Δ, srl1Δ, and tir3Δ) mutants exhibited altered mother cell size and shape. Mutants of cluster III and IV exhibited no or very small morphological defects. Cluster V (dse2Δ, egt2Δ, and sun4Δ) consisted of endoglucanase mutants with cell separation defects due to incomplete septum digestion. The cluster VI mutant cells (ecm33Δ) exhibited perturbation of apical bud growth. Cluster VII mutant cells (sag1Δ) exhibited differences in cell size and actin organization. Biochemical assays further confirmed the observed morphological defects. Further investigations based on various omics data indicated that morphological phenotyping is a complementary tool that can help with gaining a deeper understanding of the functions of mannoproteins.

    DOI PubMed

    Scopus

    1
    被引用数
    (Scopus)
  • Reliable imaging of ATP in living budding and fission yeast

    Masak Takaine, Masaru Ueno, Kenji Kitamura, Hiromi Imamura, Satoshi Yoshida

    Journal of Cell Science   132 ( 8 ) jcs230649 - jcs230649  2019年04月  [査読有り]

    担当区分:最終著者, 責任著者

    DOI PubMed

    Scopus

    15
    被引用数
    (Scopus)
  • Spindle pole body movement is affected by glucose and ammonium chloride in fission yeast

    Hiroaki Ito, Takeshi Sugawara, Soya Shinkai, Satoshi Mizukawa, Ayaka Kondo, Hisamichi Senda, Kengo Sawai, Koki Ito, Sayaka Suzuki, Masakatsu Takaine, Satoshi Yoshida, Hiromi Imamura, Kenji Kitamura, Toshinori Namba, Shin-ichi Tate, Masaru Ueno

    Biochemical and Biophysical Research Communications   511 ( 4 ) 820 - 825  2019年04月  [査読有り]

    DOI PubMed

    Scopus

    2
    被引用数
    (Scopus)
  • Kynurenine aminotransferase activity of Aro8/Aro9 engage tryptophan degradation by producing kynurenic acid in Saccharomyces cerevisiae

    Kazuto Ohashi, Romanas Chaleckis, Masak Takaine, Craig E. Wheelock, Satoshi Yoshida

    SCIENTIFIC REPORTS   7 ( 1 ) 12180  2017年09月  [査読有り]

     概要を見る

    Kynurenic acid (KA) is a tryptophan (Trp) metabolite that is synthesised in a branch of kynurenine (KYN) pathway. KYN aminotransferase (KAT) catalyses deamination of KYN, yielding KA. Although KA synthesis is evolutionarily conserved from bacteria to humans, the cellular benefits of synthesising KA are unclear. In this study, we constructed a KAT-null yeast mutant defective in KA synthesis to clarify the cellular function of KA. Amino acid sequence analysis and LC/MS quantification of KA revealed that Aro8 and Aro9 are the major KATs. KA was significantly decreased in the aro8 Delta aro9 Delta double mutant. We found that aro8 Delta aro9 Delta cells did not exhibit obvious defects in growth or oxidative stress response when proper amounts of amino acids are supplied in the media. We further found that aro8 Delta aro9 Delta cells were sensitive to excess Trp. The Trp sensitivity was not rescued by addition of KA, suggesting that Trp sensitivity is not due to the loss of KA. In conclusion, we propose that KAT activity is required for detoxification of Trp by converting it to the less toxic KA.

    DOI PubMed

    Scopus

    24
    被引用数
    (Scopus)
  • Ypk1 and Ypk2 kinases maintain Rho1 at the plasma membrane by flippase-dependent lipid remodeling after membrane stresses

    Riko Hatakeyama, Keiko Kono, Satoshi Yoshida

    JOURNAL OF CELL SCIENCE   130 ( 6 ) 1169 - 1178  2017年03月  [査読有り]

     概要を見る

    The plasma membrane (PM) is frequently challenged by mechanical stresses. In budding yeast, TORC2-Ypk1/Ypk2 kinase cascade plays a crucial role in PM stress responses by reorganizing the actin cytoskeleton via Rho1 GTPase. However, the molecular mechanism by which TORC2-Ypk1/Ypk2 regulates Rho1 is not well defined. Here, we found that Ypk1/Ypk2 maintain PM localization of Rho1 under PM stress via spatial reorganization of the lipids including phosphatidylserine. Genetic evidence suggests that this process is mediated by the Lem3-containing lipid flippase. We propose that lipid remodeling mediated by the TORC2-Ypk1/Ypk2-Lem3 axis is a backup mechanism for PM anchoring of Rho1 after PM stress-induced acute degradation of phosphatidylinositol 4,5-bisphosphate [PI(4,5) P-2], which is responsible for Rho1 localization under normal conditions. Since all the signaling molecules studied here are conserved in higher eukaryotes, our findings might represent a general mechanism to cope with PM stress.

    DOI PubMed

    Scopus

    20
    被引用数
    (Scopus)
  • Zds1/Zds2-PP2A(Cdc55) complex specifies signaling output from Rho1 GTPase

    Erin M. Jonasson, Valentina Rossio, Riko Hatakeyama, Mitsuhiro Abe, Yoshikazu Ohya, Satoshi Yoshida

    JOURNAL OF CELL BIOLOGY   212 ( 1 ) 51 - 61  2016年01月  [査読有り]

     概要を見る

    Budding yeast Rho1 guanosine triphosphatase (GTPase) plays an essential role in polarized cell growth by regulating cell wall glucan synthesis and actin organization. Upon cell wall damage, Rho1 blocks polarized cell growth and repairs the wounds by activating the cell wall integrity (CWI) Pkc1-mitogen-activated protein kinase (MAPK) pathway. A fundamental question is how active Rho1 promotes distinct signaling outputs under different conditions. Here we identified the Zds1/Zds2-protein phosphatase 2A(Cdc55) (PP2A(Cdc55)) complex as a novel Rho1 effector that regulates Rho1 signaling specificity. Zds1/Zds2-PP2A(Cdc55) promotes polarized growth and cell wall synthesis by inhibiting Rho1 GTPase-activating protein (GAP) Lrg1 but inhibits CWI pathway by stabilizing another Rho1 GAP, Sac7, suggesting that active Rho1 is biased toward cell growth over stress response. Conversely, upon cell wall damage, Pkc1-Mpk1 activity inhibits cortical PP2A(Cdc55), ensuring that Rho1 preferentially activates the CWI pathway for cell wall repair. We propose that PP2A(Cdc55) specifies Rho1 signaling output and that reciprocal antagonism between Rho1-PP2A(Cdc55) and Rho1-Pkc1 explains how only one signaling pathway is robustly activated at a time.

    DOI PubMed

    Scopus

    13
    被引用数
    (Scopus)
  • The budding yeast Polo-like kinase Cdc5 is released from the nucleus during anaphase for timely mitotic exit

    Vladimir V. Botchkarev, Valentina Rossio, Satoshi Yoshida

    CELL CYCLE   13 ( 20 ) 3260 - 3270  2014年10月  [査読有り]

     概要を見る

    Polo-like kinases are important regulators of multiple mitotic events; however, how Polo-like kinases are spatially and temporally regulated to perform their many tasks is not well understood. Here, we examined the subcellular localization of the budding yeast Polo-like kinase Cdc5 using a functional Cdc5-GFP protein expressed from the endogenous locus. In addition to the well-described localization of Cdc5 at the spindle pole bodies (SPBs) and the bud neck, we found that Cdc5-GFP accumulates in the nucleus in early mitosis but is released to the cytoplasm in late mitosis in a manner dependent on the Cdc14 phosphatase. This Cdc5 release from the nucleus is important for mitotic exit because artificial sequestration of Cdc5 in the nucleus by addition of a strong nuclear localization signal (NLS) resulted in mitotic exit defects. We identified a key cytoplasmic target of Cdc5 as Bfa1, an inhibitor of mitotic exit. Our study revealed a novel layer of Cdc5 regulation and suggests the existence of a possible coordination between Cdc5 and Cdc14 activity.

    DOI PubMed

    Scopus

    14
    被引用数
    (Scopus)
  • Comparative genetic analysis of PP2A-Cdc55 regulators in budding yeast

    Valentina Rossio, Anna Kazatskaya, Mayo Hirabayashi, Satoshi Yoshida

    CELL CYCLE   13 ( 13 ) 2073 - 2083  2014年07月  [査読有り]

     概要を見る

    Cdc55, a regulatory B subunit of the protein phosphatase 2A (PP 2A) complex, plays various functions during mitosis. Sequestration of Cdc55 from the nucleus by Zds1 and Zds2 is important for robust activation of mitotic Cdk1 and mitotic progression in budding yeast. However, Zds1-family proteins are found only in fungi but not in higher eukaryotes. In animal cells, highly conserved ENSA/ARPP-19 family proteins bind and inhibit PP 2A-B55 activity for mitotic entry.
    In this study, we compared the relative contribution of Zds1/Zds2 and ENSA-family proteins Igo1/Igo2 on Cdc55 functions in budding yeast mitosis. We confirmed that Igo1/Igo2 can inhibit Cdc55 in early mitosis, but their contribution to Cdc55 regulation is relatively minor compared with the role of Zds1/Zds2. In contrast to Zds1, which primarily localized to the sites of cell polarity and in the cytoplasm, Igo1 is localized in the nucleus, suggesting that Igo1/Igo2 inhibit Cdc55 in a manner distinct from Zds1/Zds2.
    Our analysis confirmed an evolutionarily conserved function of ENSA-family proteins in inhibiting PP 2A-Cdc55, and we propose that Zds1-dependent sequestration of PP 2A-Cdc55 from the nucleus is uniquely evolved to facilitate closed mitosis in fungal species.

    DOI PubMed

    Scopus

    7
    被引用数
    (Scopus)
  • Inhibition of Cdc42 during mitotic exit is required for cytokinesis

    Benjamin D. Atkins, Satoshi Yoshida, Koji Saito, Chi-Fang Wu, Daniel J. Lew, David Pellman

    Journal of Cell Biology   202 ( 2 ) 231 - 240  2013年07月  [査読有り]

     概要を見る

    The role of Cdc42 and its regulation during cytokinesis is not well understood. Using biochemical and imaging approaches in budding yeast, we demonstrate that Cdc42 activation peaks during the G1/S transition and during anaphase but drops during mitotic exit and cytokinesis. Cdc5/Polo kinase is an important upstream cell cycle regulator that suppresses Cdc42 activity. Failure to down-regulate Cdc42 during mitotic exit impairs the normal localization of key cytokinesis regulators- Iqg1 and Inn1-at the division site, and results in an abnormal septum. The effects of Cdc42 hyperactivation are largely mediated by the Cdc42 effector p21-activated kinase Ste20. Inhibition of Cdc42 and related Rho guanosine triphosphatases may be a general feature of cytokinesis in eukaryotes. © 2013 Atkins et al.

    DOI PubMed

    Scopus

    49
    被引用数
    (Scopus)
  • Mih1/Cdc25 is negatively regulated by Pkc1 in Saccharomyces cerevisiae.

    Kouitiro Yano, Yukifumi Uesono, Satoshi Yoshida, Akihiko Kikuchi, Jun Kashiwazaki, Issei Mabuchi, Yoshiko Kikuchi

    Genes to cells : devoted to molecular & cellular mechanisms   18 ( 6 ) 425 - 41  2013年06月  [査読有り]  [国際誌]

     概要を見る

    Mitotic cyclin-dependent kinase (CDK) is activated by Cdc25 phosphatase through dephosphorylation at the Wee1-mediated phosphorylation site. In Saccharomyces cerevisiae, regulation of Mih1 (Cdc25 homologue) remains unclear because inactivation/degradation of Swe1 (Wee1 homologue) is the main trigger for G2/M transition. By deleting all mitotic cyclins except Clb2, a strain was created where Mih1 became essential for mitotic entry at high temperatures. Using this novel assay, the essential domain of Mih1 was identified and Mih1 regulation was characterized. Mih1(3E1D) with phosphomimetic substitutions of four putative PKC target residues in Mih1 had a reduced complementation activity, whereas Mih1(4A) with those nonphosphorylatable substitutions was active. The band pattern of Mih1 by SDS-PAGE was similar to that of Mih1(4A) only after inactivation of Pkc1 in a pkc1(ts) mutant. Over-expression of GFP-tagged Mih1 or GFP-Mih1(4A) accumulated as dot-like structures in the nucleus, whereas GFP-Mih1(3E1D) was localized in the cytoplasm. Over-expression of an active form of Pkc1 excluded GFP-Mih1 from the nucleus, but had minimal effect on GFP-Mih1(4A) localization. Furthermore, addition of ectopic nuclear localization signal to the Mih1(3E1D) sequence recovered complementation activity and nuclear localization. These results suggest that Mih1 is negatively regulated by Pkc1-mediated phosphorylation, which excludes it from the nucleus under certain conditions.

    DOI PubMed

    Scopus

    8
    被引用数
    (Scopus)
  • Nuclear PP2A-Cdc55 prevents APC-Cdc20 activation during the spindle assembly checkpoint

    Valentina Rossio, Takeshi Michimoto, Takeshi Sasaki, Iwai Ohbayashi, Yoshiko Kikuchi, Satoshi Yoshida

    Journal of Cell Science   126 ( 19 ) 4396 - 4405  2013年  [査読有り]

     概要を見る

    Cdc55, a regulatory B-subunit of protein phosphatase 2A (PP2A) complex, is essential for the spindle assembly checkpoint (SAC) in budding yeast, but the regulation and molecular targets of PP2A-Cdc55 have not been clearly defined or are controversial. Here, we show that an important target of Cdc55 in the SAC is the anaphase-promoting complex (APC) coupled with Cdc20 and that APC-Cdc20 is kept inactive by dephosphorylation by nuclear PP2A-Cdc55 when spindle is damaged. By isolating a new class of Cdc55 mutants specifically defective in the SAC and by artificially manipulating nucleocytoplasmic distribution of Cdc55, we further show that nuclear Cdc55 is essential for the SAC. Because the Cdc55-binding proteins Zds1 and Zds2 inhibit both nuclear accumulation of Cdc55 and SAC activity, we propose that spatial control of PP2A by Zds1 family proteins is important for tight control of SAC and mitotic progression. © 2013. Published by The Company of Biologists Ltd.

    DOI PubMed

    Scopus

    10
    被引用数
    (Scopus)
  • Proteasomal Degradation Resolves Competition between Cell Polarization and Cellular Wound Healing

    Keiko Kono, Yasushi Saeki, Satoshi Yoshida, Keiji Tanaka, David Pellman

    CELL   150 ( 1 ) 151 - 164  2012年07月  [査読有り]

     概要を見る

    Cellular wound healing, enabling the repair of membrane damage, is ubiquitous in eukaryotes. One aspect of the wound healing response is the redirection of a polarized cytoskeleton and the secretory machinery to the damage site. Although there has been recent progress in identifying conserved proteins involved in wound healing, the mechanisms linking these components into a coherent response are not defined. Using laser damage in budding yeast, we demonstrate that local cell wall/membrane damage triggers the dispersal of proteins from the site of polarized growth, enabling their accumulation at the wound. We define a protein-kinase-C-dependent mechanism that mediates the destruction of the formin Bni1 and the exocyst component Sec3. This degradation is essential to prevent competition between the site of polarized growth and the wound. Mechanisms to overcome competition from a preexisting polarized cytoskeleton may be a general feature of effective wound healing in polarized cells.

    DOI PubMed

    Scopus

    71
    被引用数
    (Scopus)
  • A yeast GSK-3 kinase Mck1 promotes Cdc6 degradation to inhibit DNA re-replication.

    Ikui AE, Rossio V, Schroeder L, Yoshida S

    PLoS genetics   8 ( 12 ) e1003099  2012年  [査読有り]

    DOI PubMed

    Scopus

    21
    被引用数
    (Scopus)
  • Spatial regulation of Cdc55-PP2A by Zds1/Zds2 controls mitotic entry and mitotic exit in budding yeast

    Valentina Rossio, Satoshi Yoshida

    JOURNAL OF CELL BIOLOGY   193 ( 3 ) 445 - 454  2011年05月  [査読有り]

     概要を見る

    Budding yeast CDC55 encodes a regulatory B subunit of the PP2A (protein phosphatase 2A), which plays important roles in mitotic entry and mitotic exit. The spatial and temporal regulation of PP2A is poorly understood, although recent studies demonstrated that the conserved proteins Zds1 and Zds2 stoichiometrically bind to Cdc55-PP2A and regulate it in a complex manner. Zds1/Zds2 promote Cdc55-PP2A function for mitotic entry, whereas Zds1/Zds2 inhibit Cdc55-PP2A function during mitotic exit. In this paper, we propose that Zds1/Zds2 primarily control Cdc55 localization. Cortical and cytoplasmic localization of Cdc55 requires Zds1/Zds2, and Cdc55 accumulates in the nucleus in the absence of Zds1/Zds2. By genetically manipulating the nucleocytoplasmic distribution of Cdc55, we showed that Cdc55 promotes mitotic entry when in the cytoplasm. On the other hand, nuclear Cdc55 prevents mitotic exit. Our analysis defines the long-sought molecular function for the zillion different screens family proteins and reveals the importance of the regulation of PP2A localization for proper mitotic progression.

    DOI PubMed

    Scopus

    43
    被引用数
    (Scopus)
  • Mechanisms for concentrating Rho1 during cytokinesis

    Satoshi Yoshida, Sara Bartolini, David Pellman

    GENES & DEVELOPMENT   23 ( 7 ) 810 - 823  2009年04月  [査読有り]

     概要を見る

    The small GTP-binding protein, Rho1/RhoA plays a central role in cytokinetic actomyosin ring (CAR) assembly and cytokinesis. Concentration of Rho proteins at the division site is a general feature of cytokinesis, yet the mechanisms for recruiting Rho to the division site for cytokinesis remain poorly understood. We find that budding yeast utilizes two mechanisms to concentrate Rho1 at the division site. During anaphase, the primary mechanism for recruiting Rho1 is binding to its guanine nucleotide exchange factors (GEFs). GEF-dependent recruitment requires that Rho1 has the ability to pass through its GDP or unliganded state prior to being GTP-loaded. We were able to test this model by generating viable yeast lacking all identifiable Rho1 GEFs. Later, during septation and abscission, a second GEF-independent mechanism contributes to Rho1 bud neck targeting. This GEF-independent mechanism requires the Rho1 polybasic sequence that binds to acidic phospholipids, including phosphatidylinositol 4,5-bisphosphate (PIP2). This latter mechanism is functionally important because Rho1 activation or increased cellular levels of PIP2 promote cytokinesis in the absence of a contractile ring. These findings comprehensively define the targeting mechanisms of Rho1 essential for cytokinesis in yeast, and are likely to be relevant to cytokinesis in other organisms.

    DOI PubMed

    Scopus

    98
    被引用数
    (Scopus)
  • Symmetry Breaking: Scaffold Plays Matchmaker for Polarity Signaling Proteins

    Benjamin D. Atkins, Satoshi Yoshida, David Pellman

    CURRENT BIOLOGY   18 ( 24 ) R1130 - R1132  2008年12月  [査読有り]

    DOI PubMed

    Scopus

    4
    被引用数
    (Scopus)
  • Plugging the GAP between cell polarity and cell cycle

    Satoshi Yoshida, David Pellman

    EMBO REPORTS   9 ( 1 ) 39 - 41  2008年01月  [査読有り]

    DOI PubMed

    Scopus

    8
    被引用数
    (Scopus)
  • Yeast formins Bni1 and Bnr1 utilize different modes of cortical interaction during the assembly of actin cables

    Shawnna M. Buttery, Satoshi Yoshida, David Pellman

    MOLECULAR BIOLOGY OF THE CELL   18 ( 5 ) 1826 - 1838  2007年05月  [査読有り]

     概要を見る

    The budding yeast formins Bni1 and Bnr1 control the assembly of actin cables. These formins exhibit distinct patterns of localization and polymerize two different populations of cables: Bni1 in the bud and Bnr1 in the mother cell. We generated a functional Bni1-3GFP that improved the visualization of Bni1 in vivo at endogenous levels. Bni1 exists as speckles in the cytoplasm, some of which colocalize on actin cables. These Bni1 speckles display linear, retrograde-directed movements. Loss of polymerized actin or specifically actin cables abolished retrograde movement, and resulted in depletion of Bni1 speckles from the cytoplasm, with enhanced targeting of Bni1 to the bud tip. Mutations that impair the actin assembly activity of Bni1 abolished the movement of Bni1 speckles, even when actin cables were present. In contrast, Bnr1-GFP or 3GFP-Bnr1 did not detectably associate with actin cables and was not observed as cytoplasmic speckles. Finally, fluorescence recovery after photobleaching demonstrated that Bni1 was very dynamic, exchanging between polarized sites and the cytoplasm, whereas Bnr1 was confined to the bud neck and did not exchange with a cytoplasmic pool. In summary, our results indicate that formins can have distinct modes of cortical interaction during actin cable assembly.

    DOI PubMed

    Scopus

    91
    被引用数
    (Scopus)
  • Polo-like kinase Cdc5 controls the local activation of Rho1 to promote cytokinesis

    S Yoshida, K Kono, DM Lowery, S Bartolini, MB Yaffe, Y Ohya, D Pellman

    SCIENCE   313 ( 5783 ) 108 - 111  2006年07月  [査読有り]

     概要を見る

    The links between the cell cycle machinery and the cytoskeletal proteins controlling cytokinesis are poorly understood. The small guanine nucleotide triphosphate (GTP)-binding protein RhoA stimulates type II myosin contractility and formin-dependent assembly of the cytokinetic actin contractile ring. We found that budding yeast Polo-like kinase Cdc5 controls the targeting and activation of Rho1 ( RhoA) at the division site via Rho1 guanine nucleotide exchange factors. This role of Cdc5 (Polo-like kinase) in regulating Rho1 is likely to be relevant to cytokinesis and asymmetric cell division in other organisms.

    DOI PubMed

    Scopus

    122
    被引用数
    (Scopus)
  • MEN signaling: Daughter bound pole must escape her mother to be fully active

    S Yoshida, M Guillet, D Pellman

    DEVELOPMENTAL CELL   9 ( 2 ) 168 - 170  2005年08月  [査読有り]

     概要を見る

    Budding yeast divide asymmetrically and must therefore align the position of the mitotic spindle with the plane of division. The success of this process is monitored by a checkpoint-signaling mechanism. Two recent papers in Molecular Cell reveal an important new facet of this signal transduction pathway.

    DOI PubMed

    Scopus

    2
    被引用数
    (Scopus)
  • Ras recruits mitotic exit regulator Lte1 to the bud cortex in budding yeast

    S Yoshida, R Ichihashi, A Toh-e

    JOURNAL OF CELL BIOLOGY   161 ( 5 ) 889 - 897  2003年06月  [査読有り]

     概要を見る

    A Cdc25 family protein Lte1 (low temperature essential) is essential for mitotic exit at a lowered temperature and has been presumed to be a guanine nucleotide exchange factor (GEF) for a small GTPase Tem1, which is a key regulator of mitotic exit. We found that Lte1 physically associates with Ras2-GTP both in vivo and in vitro and that the Cdc25 homology domain (CHD) of Lte1 is essential for the interaction with Ras2. Furthermore, we found that the proper localization of Lte1 to the bud cortex is dependent on active Ras and that the overexpression of a derivative of Lte1 without the CHD suppresses defects in mitotic exit of a Deltalte1 mutant and a Deltaras1 Deltaras2 mutant. These results suggest that Lte1 is a downstream effector protein of Ras in mitotic exit and that the Ras GEF domain of Lte1 is not essential for mitotic exit but required for its localization.

    DOI PubMed

    Scopus

    51
    被引用数
    (Scopus)
  • Budding yeast Cdc5 phosphorylates Net1 and assists Cdc14 release from the nucleolus

    S Yoshida, A Toh-e

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   294 ( 3 ) 687 - 691  2002年06月  [査読有り]

     概要を見る

    Polo-like kinase Cdc5 and Cdc14 phosphatase are essential for mitotic exit in budding yeast. Cdc14 sequestered in the nucleolus by forming a complex with Net1, a nucleolar inhibitor of Cdc14, is activated after the release from the nucleolus and Cdc5 is essential for this release. Here we show that Cdc5 affects the phosphorylation state of Net1. Tab6 is a dominant active form of Cdc14. We found that Tab6 was released from the nucleolus of cdc5 mutant cells in a cell cycle dependent manner and that the release of Tab6 (or Cdc14) was not sufficient for the cdc5 mutant to grow at a higher temperature, Altogether, we propose that Cdc5 acts to reduce affinity between Cdc14 and Net1 and that the timing of Cdc14 release is independent of Cdc5. We also provide evidence that the critical function of Cdc5, other than Cdc14 liberation, exists in late mitotic events. (C) 2002 Elsevier Science (USA). All rights reserved.

    DOI PubMed

    Scopus

    53
    被引用数
    (Scopus)
  • Mitotic exit network controls the localization of Cdc14 to the spindle pole body in Saccharomyces cerevisiae

    S Yoshida, K Asakawa, A Toh-E

    CURRENT BIOLOGY   12 ( 11 ) 944 - 950  2002年06月  [査読有り]

     概要を見る

    Budding yeast Cdc14 phosphatase plays essential roles in mitotic exit. Cdc14 is sequestered in the nucleolus by its inhibitor Net1/Cfi1 and is only released from the nucleolus during anaphase to inactivate mitotic CDK. It is believed that the mitotic exit network (MEN) is required for the release of Cdc14 from the nucleolus because liberation of Cdc14 by net1/cfi1 mutations bypasses the essential role of the MEN. But how the MEN residing at the spindle pole body (SPB) controls the association of Cdc14 with Net1/Cfi1 in the nucleolus is not yet understood [1, 2]. We found that Cdc14-5GFP was released from the nucleolus in the MEN mutants (tem1, cdc15, dbf2, and nud1), but not in the cdc5 cells during early anaphase. The Cdc14 liberation from the nucleolus was inhibited by the Mad2 checkpoint and by the Bub2 checkpoint in a different manner when microtubule organization was disrupted. We observed Cdc14-5GFP at the SPB in addition to the nucleolus. The SPB localization of Cdc14 was significantly affected by the MEN mutations and the bub2 mutation. We conclude that Cdc14 is released from the nucleolus at the onset of anaphase in a CDC5-dependent manner and that MEN factors possibly regulate Cdc14 release from the SPB.

    DOI PubMed

    Scopus

    75
    被引用数
    (Scopus)
  • Regulation of the localization of Dbf2 and Mob1 during cell division of Saccharomyces cerevisiae

    S Yoshida, A Toh-e

    GENES & GENETIC SYSTEMS   76 ( 2 ) 141 - 147  2001年04月  [査読有り]

     概要を見る

    The mitotic exit network (MEN) governs Cdk inactivation. In budding yeast, MEN consists of the protein phosphatase Cdc14, the ras-like GTPase Tem1, protein kinases Cdc15, Cdc5, Dbf2 and Dbf2-binding protein Mob1. Tem1, Dbf2, Cdc5 and Cdc15 have been reported to be localized at the spindle pole body (SPB). Here we report changes of the localization of Dbf2 and Mob1 during cell division. Dbf2 and Mob1 localize to the SPBs in anaphase and then moves to the bud neck, just prior to actin ring assembly, consistent with their role in cytokinesis. The neck localization, but not SPB localization, of Dbf2 was inhibited by the Bub2 spindle checkpoint. Cdc14 is the downstream target of Dbf2 in Cdk inactivation, but we found that the neck localization of Dbf2 and Mob1 was dependent on the Cdc14 activity, suggesting that Dbf2 and Mob1 function in cytokinesis at the end of the mitotic signaling cascade.

  • A novel functional domain of cdc15 kinase is required for its interaction with Tem1 GTPase in Saccharomyces cerevisiae

    K Asakawa, S Yoshida, F Otake, A Toh-e

    GENETICS   157 ( 4 ) 1437 - 1450  2001年04月  [査読有り]

     概要を見る

    Exit from mitosis requires the inactivation of cyclin-dependent kinase (CDK) activity. In the budding yeast. Saccharomyces cerevisiae, a number of gene products have been identified as components of the signal transduction network regulating inactivation of CDK (called the MEN, for the mitotic exit network). Cdc15, one of such components of the MEN, is an essential protein kinase. By the two-hybrid screening, we identified Cdc15 as a binding protein of Tem1, GTPase, another essential regulator of the MEN. Coprecipitation experiments revealed that Tem1 binds to Cdc15 in vivo. By deletion analysis. we found that the Tem1-binding domain resides near the conserved kinase domain of Cdc15. The Cdc15-LF mutation. which was introduced into the Tem1-binding domain, reduced the interaction with Cdc15 and Tem1 and caused temperature-sensitive growth. The kinase activity of Cdc15 was not so much affected by the Cdc15-LF mutation. However,Cdc15-LF failed to localize to the SPB at the restrictive temperature. Our data show that the interaction with Tem1 is important for the function of Cdc15 and that Cdc15 and Tem1 function in a complex to direct the exit from mitosis.

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Misc

  • 細胞膜修復機構におけるダイナミックな極性切り替えとプロテアソームの役割

    河野 恵子, 佐伯 泰, 吉田 知史, 田中 啓二, Pellman David

    細胞工学   31 ( 10 ) 1152 - 1153  2012年09月

  • Yeast Formins Bni1 and Bnr1 Utilize Different Modes of Cortical Interaction during the Assembly of Actin Cables

    S. Buttery, S. Yoshida, D. Pellman

    MOLECULAR BIOLOGY OF THE CELL   17  2006年

    研究発表ペーパー・要旨(国際会議)  

  • Regulation of budding yeast formin Bni1

    S Yoshida, D Pellman

    MOLECULAR BIOLOGY OF THE CELL   15   266A - 266A  2004年11月

    研究発表ペーパー・要旨(国際会議)  

  • Ras recruits mitotic exit regulator Lte1 to the bud cortex in budding yeast.

    S Yoshida, R Ichihashi, A Toh-e

    YEAST   20   S258 - S258  2003年07月

    研究発表ペーパー・要旨(国際会議)  

  • Spindle checkpoint control in budding yeast

    A Toh-e, K Asakawa, S Yoshida

    JOURNAL OF MICROBIOLOGY   39 ( 1 ) 1 - 10  2001年03月

    書評論文,書評,文献紹介等  

  • M期終了機構はどこまでわかったか (特集 細胞周期--急速に解明される分子メカニズムとその応用)

    浅川 和秀, 吉田 知史, 東江 昭夫

    細胞工学   19 ( 4 ) 556 - 563  2000年04月

    CiNii

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受賞

  • 酵母コンソーシアムフェロー

    2019年10月   大隅基礎科学創成財団  

    受賞者: 吉田知史

  • 第91回大会BP賞

    2019年09月   日本遺伝学会  

    受賞者: 吉田知史

  • 会長賞

    2018年09月   酵母遺伝学フォーラム  

    受賞者: 吉田知史

  • New Investigator Award

    2010年07月   Massachusetts Life Science Center  

    受賞者: 吉田知史

  • Medical Foundation Fellowship

    2007年07月   Charles King Trust  

    受賞者: 吉田知史

  • Young Scientists' prize

    2006年06月   国際生化学・分子生物学連合  

    受賞者: 吉田知史

  • 海外特別研究員

    2005年04月   日本学術振興会  

    受賞者: 吉田知史

  • 特別研究員 (DC2)

    2001年04月   日本学術振興会  

    受賞者: 吉田知史

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共同研究・競争的資金等の研究課題

  • 細胞外小胞生成に必要な遺伝子の網羅的同定とその解析

    科学技術振興機構  さきがけ

    研究期間:

    2018年
    -
    2022年
     

    吉田知史

  • 環境変化に応答してRho1GTPaseがシグナルアウトプットを変化させる仕組み

    日本学術振興会  科研費 基盤B

    研究期間:

    2016年
    -
    2019年
     

    吉田知史

  • RhoのGTPase活性を標的とした抗がん剤開発の合理性を明らかにする

    武田科学振興財団  医学系研究奨励

    研究期間:

    2016年
    -
     
     

    吉田知史

学内研究費(特定課題)

  • 発がん遺伝子RhoのGTPase活性はなぜ必須なのか?

    2020年  

     概要を見る

    Rho GTPaseはGTPと結合することによって活性化し何種類もの下流の標的タンパク質を活性化する。しかし細胞内外からの刺激により活性化したRhoは多種類の標的分子を全て均等に活性化するのではなく、刺激(入力シグナル)の種類に応じて特定の標的(出力シグナル)のみを選択的に活性化する。入力シグナルはRhoを活性化すると同時に出力シグナル経路を規定していると予想されるがその分子機構はよくわかっていない。本研究ではGTPを加水分解できないRho1-G19V, Rho1-Q68L変異体を作成し酵母細胞内でその機能を解析し、Rho1-G19V, Rho1-Q68Lは試験管内では下流の標的因子を強く活性化するにもかかわらず細胞内では機能できないことを明らかにした。この発見は主要な発癌遺伝子であるRhoA GTPaseの阻害剤の開発が可能であることを証明するものであり,RhoAのGTPase活性阻害薬の有効性が実証できれば新しい癌治療法へのアプローチとして大きな貢献となることが期待される。

  • Dissecting the roles of ATP homeostasis in cellular senescence

    2019年  

     概要を見る

    細胞の活性状態と細胞の老化には密接な関連があると予想されているが確定した根拠はない。本研究では細胞内エネルギーをATP濃度を指標とすることで計測し細胞がストレス応答や老化の過程でATP濃度をどのように保持するのか?またATP濃度に異常が生じた際に細胞にどのような不都合が起こるのかを検証した。これまでに我々は生きている個々の細胞内でのATP濃度を定量的に測定する系の開発に成功した (Takaine et al., J Cell Sci. 2019, Ito et al., BBRC. 2019)。この系を発展させ老化を含む様々なコンディションでのATP濃度変化を可視化へと挑戦している。

  • 細胞内エネルギー状態を可視化し、代謝状態と老化および疾患との関連を明らかにする

    2018年  

     概要を見る

    採択者は2018年に群馬大学生体調節研究所から早稲田大学国際教養学部へと異動した。本特定課題研究によるサポートにより早稲田大学で新しく研究室を立ち上げるのに必須な器具類の購入が可能となった。研究室のセットアップは少しずつではあるが進んでおり2018年度には本研究課題に関連した「細胞内エネルギーATPの可視化」に関する2報の共同研究論文(Ito et al., BBRC. 2019, および Takaine et al., J. Cell Sci. 2019)を出版することができた。さらに本研究課題を発展させた新規プロジェクトの提案「細胞外微粒子に必須な遺伝子群の網羅的同定」が科学技術振興機構のさきがけに採択された。早稲田大学で研究を発展させていくための基盤が徐々に整いつつあると考えている。

 

現在担当している科目

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