FY2021 Annual Report

Membranology Unit
Assistant Professor Keiko Kono

Abstract

In FY2021, we continuously investigated the molecular mechanisms and physiological consequences of plasma membrane damage using budding yeast and normal human fibroblasts. We reported that the clathrin-flippase-exocyst regulatory axis is involved in plasma membrane repair factors delivery in budding yeast (Yamazaki and Kono, BBRC, 2021). The first author Mr. Yuta Yamazaki, a Ph.D. student at OIST, published this paper as early as six months after his joining the lab. We also investigated the time-resolved transcriptomic and proteomic changes during plasma membrane damage-induced senescence in normal human fibroblasts. We are preparing three manuscripts on this topic. We hosted two rotation student, and welcomed two new Ph.D. students.

1. Staff

  • Dr. Yohsuke Moriyama, Science and Technology Associate
  • Dr. Shinju Sugiyama, Postdoc
  • Dr. Nurhanani Binti Razali, Postdoc
  • Mr. Hunter Barbee, Ph.D. Student
  • Ms. Yatzu Chiu, Ph.D. Student
  • Mr. Kojiro Suda, Ph.D. Student
  • Mr. Jan Grašič, Ph.D. Student
  • Mr. Yuta Yamazaki, Ph.D. Student
  • Ms. Kamila Krystyna Kozik, Ph.D. Student
  • Ms. Tara Helmi Turkki, Rotation Student
  • Ms. Hitomi Ohtaki, Research Unit Administrator

Co-supervision

  • Ms. Aisulu Maipas (Ph.D. Student@ Yamamoto Unit)
  • Ms. Sara Abdelaal (Ph.D. Student@ Terenzio Unit)
  • Mr. Masato Hirota (Ph.D. Student@ Ishikawa Unit)

2. Collaborations

2.1  Immune-cell-like functions of epitherial cells

  • Type of collaboration: Joint research
  • Researchers:
    • Dr. Takeshi Maruyama, TWIns, Waseda University

2.2 Plasma membrane damage response in Naked Mole Rat

  • Type of collaboration: Joint research
  • Researchers:
    • Dr. Kyoko Miura, Kumamoto University

2.2 Plasma membrane damage response in laboratory yeast and industrial yeast for brewing

  • Type of collaboration: Joint research
  • Researchers:
    • Dr. Nozomu Kamada, SUNTORY
    • Dr. Hiroshi Kawashima, SUNTORY
    • Dr. Atsushi Fujita, SUNTORY
    • Dr. Fumihiko Omura, SUNTORY
    • Dr. Yohei Fujita, SUNTORY

3. Activities and Findings

Clathrin-mediated trafficking of phospholipid flippases is required for local plasma membrane/cell wall damage repair in budding yeast

Plasma membrane damage and repair frequently happen in cells. A critical process underlying plasma membrane repair is to redirect repair factors, such as protein kinase C and the exocyst complex, from the polarized site to the damage site. However, the mechanism underlying the repair factor delivery to the damage site remains unknown. Here, we demonstrate that clathrin-mediated trafficking of repair factors is involved in plasma membrane/cell wall repair in budding yeast. Using laser-induced plasma membrane/cell wall damage assay, we identified phospholipid flippases, Lem3-Dnf1/Dnf2 and Cdc50-Drs2, as essential clathrin cargos for plasma membrane/cell wall repair. We found that flippase impairment significantly compromised the recruitment of exocyst Exo70 to the damage site. In contrast, the recruitment of protein kinase C (Pkc1) was only mildly compromised. Taken together, clathrin-mediated trafficking of the phospholipid flippases is critical for the recruitment of exocyst to the damage site. Mechanisms to redirect exocyst via the clathrin and flippase-mediated pathways may be a general feature of effective plasma membrane repair in polarized cells.

4. Publications

4.1 Journals

  1. Yamazaki Y, Kono K “Clathrin-mediated trafficking of phospholipid flippases is required for local plasma membrane/cell wall damage repair in budding yeast” Biochemical and Biophysical Research Communications, doi: 10.1016/j.bbrc.2022.03.129 (2021).

4.2 Books and other one-time publications

Nothing to report

4.3 Oral and Poster Presentations

Oral presentation (International)

  1. Moriyama Y, Razali NB, Chiu Y, Suda K and Kono K. “Plasma membrane damage limits replicative lifespan in budding yeast and induces premature senescence in normal human fibroblasts” (2021 Dec) The 6th international Cell Senescence Association Conference, Osaka. A talk selected from abstract.
  2. Kono K. “Plasma membrane damage induces cellular senescence” (2021 Sep) Ohio University Biomedical Engineering Seminar. An invited lecture.

 

Oral presentation (Domestic)

  1. Kono K. “Plasma membrane damage-induced senescent cells promote wound healing via paracrine signaling” (2022 Mar), Riken, Online
  2. Kono K. “Wounding alters yeast’s fate” (2022 Feb) The 200th Yeast Cell colloquium, Online, Symposium
  3. Kono K. “Plasma membrane damage limits replicative lifespan in budding yeast and human fibroblasts” (2021 Nov) The 94th Annual Meeting of the Japanese Biochemical Society, Hybrid, Symposium
  4. Kono K. “Cortical tension and chromosome segregation” (2021 Sep) The 93rd Annual Meeting of the Genetic Society of Japan, Online, Workshop
  5. Kono K. “Down-regulating cortical tension ensures timely chromosome segregation.” (2021 June) The 73rd Annual Meeting of the Japan Society for Cell Biology, Online, Symposium
  6. Sugiyama S, Kono K. "Spatial regulation of ER-PM contact site proteins at polarity sites” (2021 Jul) The 7th meeting of Associations for Yeast Young Researchers Online
  7. Sugiyama S, Kono K. "Exclusion of ER-PM contact site proteins from polarity sites in budding yeasts." (2021 Sep) The 54th annual meeting of Yeast Genetic Society of Japan Online
  8. Yamazaki Y, Kono K. "A role of clathrin in plasma membrane repair"  (2021 Sep) The 54th annual meeting of Yeast Genetic Society of Japan Online

Poster Presentation​

  1. Razali N, Moriyama Y, Chiu Y, Kono K. "Plasma membrane damage induces wound-healing SASPs in normal human fibroblasts." (2021 Dec) The 6th International Cell Senescence Association Conference Osaka International Convention Center, Osaka
  2. Chiu Y, Moriyama Y, Razali, N, Kono K. (2021 Dec) “The miRNA Signature Associated with Plasma Membrane Damage-dependent Senescence” The 6th International Cell Senescence Association Conference. Osaka International Convention Center, Osaka

5. Intellectual Property Rights and Other Specific Achievements

Nothing to report

6. Meetings and Events

Nothing to report

7. Other

Nothing to report.