Takanari Inoue

Takanari Inoue, Ph.D.

Professor

443-287-7668 (office), 443-287-7669 (lab)

Department of Cell Biology
Johns Hopkins University School of Medicine
855 N. Wolfe Street, 476 Rangos
Baltimore, MD 21205

Website

Academic Titles

Professor

Research Topic

Directed cell migration; tumor metastasis; primary cilia; synthetic chemical biology; technology development

Research Topic: Synthetic Cell Biology

Our research focuses on synthetic cell biology to dissect intricate signaling networks. In particular, we investigate positive-feedback mechanisms underlying the initiation of neutrophil chemotaxis (known as a symmetry breaking process), as well as spatio-temporally compartmentalized Ras signaling. In parallel, our lab also tries to understand how cell morphology affects biochemical functions in cells. Ultimately, we will generate completely orthogonal nano-machinery in cells that can achieve existing, and even novel, cellular functions.

Figure 1: Engineering cellular morphology. Time series of confocal fluorescence images of NIH3T3 fibroblasts experiencing striking morphological changes upon rapid activation of small GTPases such as Rac, Cdc42, and RhoA.

Figure 2: Inducible cell migration. Time series of confocal dual-color fluorescence images of differentiated HL-60 cells in which PI3K or Rac is rapidly and inducibly activated without activating any molecules upstream. Such synthetic activation of PI3K, but not Rac, led to cell polarization and following migration. Yellow: PI3K or Rac activated cells, Blue: Nuclei.

Figure 3: Tools to manipulate protein activity at specific subcellular compartments (a) A principle diagram for inducible recruitment of signaling molecules (green circles). c, cytoplasm, o, organelle. (b) Confocal fluorescence images of HeLa cells showing rapid translocation of cytoplasmic proteins to specific organelles upon 5 M iRap addition for 5 minutes. Scale bar indicates 20 m.

Biosketch

Lab Members

Name Classification Email Phone
Abhijit Deb Roy, Ph.D. Postdoctoral Fellow adebroy1@jhmi.edu 443-287-7669
Cristian Saez Gonzalez Student csaezgo1@jhu.edu
Di (Helen) Wu Student dwu32@jhu.edu 443-287-7669
Edward Cai Student ecai1@jhu.edu 443-287-7669
Robert DeRose Research Specialist jderose1@jhmi.edu 443-287-7669
Yuta Nihongaki, Ph.D. Postdoctoral Fellow ynihong1@jhmi.edu 443-287-7669

Publications

  • Wolfe K., Kamata R., Coutinho K., Inoue T. and Sasaki A.T. “Metabolic Compartmentalization at the Leading Edge of Metastatic Cancer Cells” Frontiers in Oncology (in press)
  • Kim AK, Wu HD, Inoue T, “Rational Design of a Protein Kinase A Nuclear-cytosol Translocation Reporter” Scientific Reports 2020:10:9365
  • Wu HD, Kikuchi M, Dagliyan O, Aragaki AK, Nakamura H, Dokholyan NV, Umehara T, Inoue T, “Rational design and implementation of a chemically inducible hetero-trimerization system” Nature Methods 17, 928–936 (2020). Note: Research Highlight in Nature Chemical Biology, “Three’s company”

  • Georgess D, Padmanaban V, Sirka OK, Coutinho K, Choi A, Frid G, Neumann NM, Inoue T, Ewald AJ. Twist1-induced epithelial dissemination requires Prkd1signaling. Cancer Res, 2020 Jan 15, 80 (2), 204-218. 

  • Nakamura H, DeRose R, Inoue T. “Harnessing biomolecular condensates in living cells” Journal of Biochemistry (in press, 2019)

  • Miao Y, Battacharya S, Banerjee T, Abudaker-Sharif B, Long Y, Inoue T, Iglesias P and Devreotes P. (2019). Wave patterns organize cellular protrusions and control cortical dynamics. Molecular Systems Biology 15(3). DOI 10.15252/msb.20188585| Published online 11.03.2019.

  • Cuei-Ling Wang, Yu-Chen Chang, Ya-Chu Chang, Shi-Rong Hong, Chun-Yu Lin, Ning Hsu, Hsiao-Chi Cheng, Yueh-Chen Chiang, Wei-En Huang, Nathan C. Shaner, Rajat Rohatgi, Inoue T, Yu-Chun Lin. (2018).“Spatiotemporal manipulation of ciliary glutamylation reveals its roles in intraciliary trafficking and Hedgehog signaling.”Nature Communications 2018:9:1732:1-13

  • Naoto Takada, Tomoki Naito, Inoue T, Kazuhisa Nakayama, Hiroyuki Takatsu, Hye-Won Shin. “Enhanced phospholipid flipping by P4-ATPase promotes membrane deformation.” 2018. EMBO Reports (in press). 

  • Nakamura H, Lee AA, Afshar AS, Lin YC, Tanigawa M, Suarez A, Razavi S, DeRose R, Bobb D, Hong W, Gabelli SB, Goutsias J, Inoue T. 2017. Intracellular production of hydrogels and synthetic RNA granules by multivalent molecular interactions. Nature Materials (In press, doi:10.1038/nmat5006)

  • Miao Y, Bhattacharya S, Edwards M, Cai H, Inoue T, Iglesias P, Devreotes PN. 2017.  Altering the threshold of an excitable signal transduction network changes cell migratory modes.  Nat Cell Biol 19:329-340.

  • Phua SC, Chiba S, Suzuki M, Su E, Roberson EC, Pusapati GV, Setou M, Rohatgi R, Reiter JF, Ikegami K, Inoue T. 2017. Dynamic Remodeling of Membrane Composition Drives Cell Cycle through Primary Cilia Excision. Cell 2017;168:264-279

  • Niu J., Johny M.B., Dick I.E., Inoue T. 2016. Following Optogenetic Dimerizers and Quantitative Prospects. Biophysical Journal 111:1132–1140

  • Gaus K., Inoue T. 2016. New Biological Frontiers Illuminated by Molecular Sensors and Actuators. Biophysical journal. 2016; 111(6):E01-2.

  • Meier EL, Razavi S, Inoue T, Goley ED. A novel membrane anchor for FtsZ is linked to cell wall hydrolysis in Caulobacter crescentus. Mol Microbiol. 2016 Jul;101(2):265-80.

  • Gaus K., Inoue T. 2016. New Biological Frontiers Illuminated by Molecular Sensors and Actuators. Biophysical Journal. 2016;111(6):E01–E02

  • Coutinho K, Inoue T. 2016. Deconstructing and constructing innate immune functions using molecular sensors and actuators. Proc. SPIE 9871, Sensing and Analysis Technologies for Biomedical and Cognitive Applications 2016, 987102 (May 19, 2016); doi:10.1117/12.2225185 

  • Antonczak A.K., Mullee L., Wang Y., Comartin D., Inoue T., Pelletier L. and Morrison C.G. 2016. Opposing effects of pericentrin and microcephalin on the pericentriolar material regulate CHK1 activation in the DNA damage response.  30:2003-10. 

  • Kim AK, DeRose R, Ueno T, Lin B, Komatsu T, Nakamura H, Inoue T. Toward total synthesis of cell function: Reconstituting cell dynamics with synthetic biology. Sci Signal. 2016 Feb 9;9(414). Review.

  • Phua SC, Lin YC, Inoue T. An intelligent nano-antenna: Primary cilium harnesses TRP channels to decode polymodal stimuli. Cell Calcium. 2015 Oct;58(4):415-22. Review.

  • Miyamoto T, Rho E, Inoue T. Deconvoluting AMPK dynamics. Oncotarget. 2015 Oct 13;6(31):30431-2.

  • Garcia-Gonzalo FR, Phua SC, Roberson EC, Garcia G 3rd, Abedin M, Schurmans S, Inoue T, Reiter JF. Phosphoinositides regulate ciliary protein trafficking to modulate hedgedog signaling. Dev Cell. 2015 Aug 24;34(4):400-9.

  • Nguyen H-N, Yang J-M, Miyamoto T, Itoh K, Rho E, Zhang Q, Inoue T, Devreotes PN, Sesaki H, and Iijima M. 2015. Opening the conformation is a master switch for the dual localization and phosphatase activity of PTEN.  Sci Rep Jul 28;5:12600. doi: 10.1038/srep12600. PMCID: PMC4517176.

  • Lin B., Yin T., Wu Y.I., Inoue T.* and Levchenko A.* “Interplay between chemotaxis and contact inhibition of locomotion determines exploratory cell migration” (Accepted at Nature Communications)

  • Miyamoto T., Rho E, Sample V., Akano H., Magari M., Ueno T., Chen M., Tokumitsu H., Zhang J., and Inoue T.* “Compartmentalized AMPK Signaling Illuminated by Genetically Encoded Molecular Sensors and Actuators” (Accepted at Cell Reports)

  • Onuma H, Komatsu T, Arita M, Hanaoka K, Ueno T, Terai T, Nagano T, Inoue T.  2014.  Rapidly rendering cells phagocytic through a cell surface display technique and concurrent Rac activation.  Sci Signal.  Jul 15;7(334):rs4. doi: 10.1126/scisignal.2005123.

  • Suarez A., Ueno T., Huebner R., McCaffery J.M., and Inoue T.* “Bin/Amphiphysin/Rvs (BAR) family members bend membranes in cells” Scientific Reports 4, 4693 (2014)

  • Kobayashi T., Kim S., Lin Y.C., Inoue T., and Dynlacht B.D. “CP110-interacting proteins, Talpid3 and Cep290, play overlapping and distinct roles in cilia assembly.” Journal of Cell Biology 204, 215 (2014) 

  • Su S., Phua S.C., DeRose R., Chiba S., Narita K., Kalugin P.N., Katada T., Kontani K., Takeda S. and Inoue T. “Genetically encoded calcium indicator illuminates calcium dynamics in cilia.” Nature Methods 10, 1105 (2013)

  • Thevathasan, J.V., Tan E., Hui Z., Lin Y.C., Li Y., Inoue T. and Fivaz M. “The small GTPase HRas shapes local PI3K signals through positive feedback and regulates persistent membrane extension in migrating fibroblasts.” Molecular Biology of the Cell 24, 2228 (2013)

  • Lin Y.C., Niewiadomski P., Lin B., Nakamura H., Phua S.C., Jiao J., Levchenko A., Inoue T., Rohatgi T., and Inoue T. “Chemically-inducible diffusion trap reveals molecular sieve-like barrier at primary cilia“ Nature Chemical Biology 9, 437 (2013)

  • Lin YC, Liu T-Y, Razavi S. and Inoue T. “Rapidly Reversible Manipulation of Molecular Activities Using Dual Chemical Dimerizers” Angewandte Chemie 52, 6450 (2013)

  • Thevathasan, J.V., Tan E., Hui Z., Lin Y.C., Li Y., Inoue T. and Fivaz M. “Local positive feedback from PI3K to Ras drives cell polarization and migration” Molecular Biology of the Cell (doi:10.1091/mbc.E12-12-0905)

  • Miyamoto T., DeRose R., Suarez A., Ueno T., Chen M., Sun T.-p., Wolfgang M.J., Mukherjee C., Meyers D. and Inoue T. “Generation of Intracellular Logic Gates with Two Orthogonal Chemically Inducible Systems” Nature Chemical Biology (Accepted 2012)

  • Umeda N., Ueno T., Pohlmeyer C., Nagano T. and Inoue T. “A photocleavable rapamycin conjugate for spatiotemporal control of small GTPase activity” Journal of American Chemical Society 133(1), 12-14 (2011) Note: “Scientist use light to move molecules within living cells” Science News article in ScienceDaily

  • Ueno T., Falkenburger B.H., Pohlmeyer C., and Inoue T. “Triggering Actin Comets Versus Membrane Ruffles: Distinctive Effects of Phosphoinositides on Actin Reorganization” ScienceSignaling 4(203), ra87 (2011) (Cover Article)

  • Komatsu T., Kukelyansky I., McCaffery J.M., Ueno T., Varela L.C. and Inoue T. “Organelle-Specific, Rapid Induction of Molecular Activities and Membrane Tethering” Nature Methods 7, 206-208 (2010) Note: “Hopkins researchers put proteins right where they want them” Breaking News article in Genetic Engineering and Biotechnology News

  • Rahdar M., Inoue T., Meyer T., Zhang J., Vazquez F., and Devreotes P.N. “A phosphorylation-dependent intramolecular interaction regulates the membrane association and activity of the tumor suppressor PTEN” Proc. Natl. Acad. Sci. U S A. 106(2):480-5 (2009)

  • Inoue T. and Meyer T. “Synthetic activation of endogenous PI3K and Rac identifies an AND-gate switch for cell polarization and migration” PLoS ONE 3(8), e3068 (2008)

  • Fivaz M., Bandara S., Inoue T. and Meyer T. "Robust neuronal symmetry breaking by Ras-triggered local positive feedback" Current Biology 18, 44-50 (2008)

  • Heo WD, Inoue T, Park WS, Kim ML, Park BO, Wandless TJ, and Meyer T. “PI(3,4,5)P3 and PI(4,5)P2 lipids target Ras, Rho, Arf and Rab GTPases to the plasma membrane” Science 314, 1458-1461 (2006)

  • Suh BC*, Inoue T*, Meyer T, and Hille B. “Rapid chemically-induced changes of PtdIns(4,5)P2 gate KCNQ ion channels” Science 314, 1454-1457 (2006) (*Contributed Equally) Note: “Perspectives” (Science 314, 1402-1403 (2006)), “Editor’s Choice” (Science STKE 364, tw410 (2006)), “Spotlight” (ACS Chem. Biol. 1, 608 (2006)), “Research Highlights” (Nature Methods 4, 7 (2007))

  • Inoue T, Heo WD, Grimley JS, Wandless TJ, and Meyer T. “Inducible translocation strategies to rapidly activate and inhibit small GTPase signaling pathways” Nature Methods 2, 415-418 (2005)

  • Inoue T, Kikuchi K, Hirose K, Iino M, and Nagano T. “Spatiotemporal Laser Inactivation of Inositol 1,4,5-Trisphosphate Receptors Using Synthetic Small-molecule Probes” Chem. Biol. 10, 503-509 (2003) Note: “Cover art”

  • Inoue T, Kikuchi K, Hirose K, Iino M, and Nagano T. “Small molecule-based laser inactivation of inositol 1,4,5-trisphosphate receptor” Chem. Biol. 8, 9-15 (2001)