Devising new electron microscopy approach to explore the morphology of chromatin in cell nucleus
| Project/Area Number |
18K19301
|
|---|
| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
|
| Allocation Type | Multi-year Fund |
|---|
| Review Section |
Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
|
|---|
| Research Institution | Hiroshima University |
|---|
Principal Investigator |
Tate Shin-ichi 広島大学, 統合生命科学研究科(理), 教授 (20216998)
|
|---|
| Project Period (FY) |
2018-06-29 – 2020-03-31
|
| Project Status |
Completed (Fiscal Year 2019)
|
| Budget Amount *help |
¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
Fiscal Year 2019: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2018: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
|
|---|
| Keywords | FIB-SEM / クロマチン構造 / 分裂酵母 / 電子顕微鏡 / APEX / クロマチン構造動態 / 導電染色技術 |
|---|
| Outline of Final Research Achievements |
We have successfully observed the three-dimensional (3D) electron microscopic image of chromatin in cell-nucleus with focused ion-beam scanning electron microscopy (FIB-SEM). This gives the first example for describing the whole chromatin 3D structure within cell-nucleus at a single-cell level.
We have established the 3D structure analysis of the whole chromatin within cell nucleus. The fission yeast cells that harbor histone H2B-APEX2 fusion protein enabled the diaminobenzidine (DAB) coating around the chromatin fibers inside the cell nucleus, thus allowed the osmium-ion staining to give high contrast EM image of the chromatin within the cell nucleus. The numerical processing software devised in this research achieved very efficient reconstruction of the 3D EM image of chromatins from a series of SEM images from FIB-SEM. This EM technology will pave ways in the study of gene regulation from the whole chromatin 3D structural points of view.
|
| Academic Significance and Societal Importance of the Research Achievements |
細胞の遺伝情報制御機構の解明は，ガンなど疾病発症機構や細胞の分化制御機構の理解につながる細胞生物学の中心的なテーマである．分子生物学は，DNAとタンパク質の局所的な相互作用を明らかにしてきた．一方で，最近の研究は，核内で遺伝情報を保持する「クロマチン」構造が，局所の分子間相互作用を調節する機構の存在を示唆している．しかし，核内クロマチン全体の立体構造を観測する技術が無いため研究されていない．私達は核内クロマチンの3次元構造を明らかにする電子顕微鏡技術を構築した．この技術により，クロマチン全体の構造変化による遺伝情報制御機構を解明する新たな細胞生物学研究が可能になる．
|
Report
(3 results)
Research Products
(20 results)
-
-
[Journal Article] DOPG small unilamellar vesicles function as nano-carriers targeting the clustered lectin-like oxidized LDL receptor (LOX-1) on the cell surface2019
Author(s)
Ohta,T., Yamada,R., Fujita,S., Takahata,T., Shiba,K., Machida,S., and Tate,S.
-
Journal Title
J. Drug Delivery Sci. and Tech.
Volume: 51
Pages: 327-336
DOI
Related Report
Peer Reviewed
-
[Journal Article] Spindle pole body movement is affected by glucose and ammonium chloride in fission yeast2019
Author(s)
Ito H, Sugawara T, Shinkai S, Mizukawa S, Kondo A, Senda H, Sawai K, Ito K, Suzuki S, Takaine M, Yoshida S, Imamura H, Kitamura K, Namba T, Tate SI, Ueno M
-
Journal Title
Biochem Biophys Res Commun
Volume: 511
Issue: 4
Pages: 820-825
DOI
Related Report
Peer Reviewed
-
[Journal Article] Backbone and side-chain chemical shift assignments of full-length, apo, human Pin1, a phosphorprotein regulator with interdomain allostery2019
Author(s)
Born,A., Nichols,P.J., Henen,M.A., Chi,C.N., Strotz,D., Bayer,P., Tate,S., Peng,J.W., Vogeli,B.
-
Journal Title
Biomolecular NMR Assignments
Volume: 13
Issue: 1
Pages: 85-89
DOI
Related Report
Peer Reviewed / Int'l Joint Research
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
