A collection of yeast cellular electron cryotomography data

被引:5
作者
Gan, Lu [1 ,2 ]
Ng, Cai Tong [1 ,2 ]
Chen, Chen [1 ,2 ]
Cai, Shujun [1 ,2 ]
机构
[1] Natl Univ Singapore, Dept Biol Sci, 14 Sci Dr 4, Singapore 117543, Singapore
[2] Natl Univ Singapore, Ctr BioImaging Sci, 14 Sci Dr 4, Singapore 117543, Singapore
来源
GIGASCIENCE | 2019年 / 8卷 / 06期
关键词
yeast; chromatin; nucleus; cryo-ET; cryo-EM; template matching; mining; CRYOELECTRON TOMOGRAPHY; CRYOMICROSCOPY; ALIGNMENT; CELLS;
D O I
10.1093/gigascience/giz077
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: Cells are powered by a large set of macromolecular complexes, which work together in a crowded environment. The in situ mechanisms of these complexes are unclear because their 3D distribution, organization, and interactions are largely unknown. Electron cryotomography (cryo-ET) can address these knowledge gaps because it produces cryotomograms-3D images that reveal biological structure at similar to 4-nm resolution. Cryo-ET uses no fixation, dehydration, staining, or plastic embedment, so cellular features are visualized in a life-like, frozen-hydrated state. To study chromatin and mitotic machinery in situ, we subjected yeast cells to genetic and chemical perturbations, cryosectioned them, and then imaged the cells by cryo-ET. Findings: Here we share > 1,000 cryo-ET raw datasets of cryosectioned budding yeast Saccharomyces cerevisiae collected as part of previously published studies. These data will be valuable to cell biologists who are interested in the nanoscale organization of yeasts and of eukaryotic cells in general. All the unpublished tilt series and a subset of corresponding cryotomograms have been deposited in the EMPIAR resource for the community to use freely. To improve tilt series discoverability, we have uploaded metadata and preliminary notes to publicly accessible Google Sheets, EMPIAR, and GigaDB. Conclusions: Cellular cryo-ET data can be mined to obtain new cell-biological, structural, and 3D statistical insights in situ. These data contain structures not visible in traditional electron-microscopy data. Template matching and subtomogram averaging of known macromolecular complexes can reveal their 3D distributions and low-resolution structures. Furthermore, these data can serve as testbeds for high-throughput image-analysis pipelines, as training sets for feature-recognition software, for feasibility analysis when planning new structural-cell-biology projects, and as practice data for students.
引用
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页数:9
相关论文
共 55 条
[31]  
Marini G, 2018, BIORXIV, DOI DOI 10.1101/468454
[32]   Dual-axis tomography: An approach with alignment methods that preserve resolution [J].
Mastronarde, DN .
JOURNAL OF STRUCTURAL BIOLOGY, 1997, 120 (03) :343-352
[33]   Automated electron microscope tomography using robust prediction of specimen movements [J].
Mastronarde, DN .
JOURNAL OF STRUCTURAL BIOLOGY, 2005, 152 (01) :36-51
[34]   Experimental observation of the improvement in MTF from backthinning a CMOS direct electron detector [J].
McMullan, G. ;
Faruqi, A. R. ;
Henderson, R. ;
Guerrini, N. ;
Turchetta, R. ;
Jacobs, A. ;
van Hoften, G. .
ULTRAMICROSCOPY, 2009, 109 (09) :1144-1147
[35]   Characterization of a direct detection device imaging camera for transmission electron microscopy [J].
Milazzo, Anna-Clare ;
Moldovan, Grigore ;
Lanman, Jason ;
Jin, Liang ;
Bouwer, James C. ;
Klienfelder, Stuart ;
Peltier, Steven T. ;
Ellisman, Mark H. ;
Kirkland, Angus I. ;
Xuong, Nguyen-Huu .
ULTRAMICROSCOPY, 2010, 110 (07) :741-744
[36]   Electron cryotomography analysis of Dam1C/DASH at the kinetochore-spindle interface in situ [J].
Ng, Cai Tong ;
Deng, Li ;
Chen, Chen ;
Lim, Hong Hwa ;
Shi, Jian ;
Surana, Uttam ;
Gan, Lu .
JOURNAL OF CELL BIOLOGY, 2019, 218 (02) :455-473
[37]   The molecular architecture of axonemes revealed by cryoelectron tomography [J].
Nicastro, Daniela ;
Schwartz, Cindi ;
Pierson, Jason ;
Gaudette, Richard ;
Porter, Mary E. ;
McIntosh, J. Richard .
SCIENCE, 2006, 313 (5789) :944-948
[38]   Automated batch fiducial-less tilt-series alignment in Appion using Protomo [J].
Noble, Alex J. ;
Stagg, Scott M. .
JOURNAL OF STRUCTURAL BIOLOGY, 2015, 192 (02) :270-278
[39]  
Nuske E, 2018, BIORXIV, DOI [10.1101/467829, DOI 10.1101/467829]
[40]  
O'Toole E, 2018, ETOMO TUTORIAL IMOD