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Chromosomes are the first and final frontier of structural cell biology. They contain the instructions of life, which must be read, copied, and passed on to daughter cells. These activities must be done accurately by each of the trillions of cells in the human body. Mistakes can lead to many diseases, including cancers. How do cells access and use the information stored in chromosomes? How do they fold up the fundamental subunits called nucleosomes and then transport them into daughter cells? The answers to many of these fundamental questions will require knowledge of the organization, positions, and distributions of the subunits of chromosomes and their interaction partners, all within their native intracellular context.
The DNA and proteins that make up chromosomes are measured in nanometers, but they reside inside cells that are measured in micrometers. To map out the nanoscale organization of chromosomes inside their native cellular context, we use state-of-the-art electron tomography to generate 3-D images called “tomograms”. To minimize the fixation, dehydration, extraction, and staining artifacts associated with conventional electron microscopy (EM), we prepare and image our cells in a life-like frozen-hydrated state. This combination of 3-D nanoscale imaging and cryogenic sample handling is called cryo-electron tomography (cryo-ET). All our work has been done at the NUS Center for Bioimaging Sciences, which is equipped with top-of-the-line instruments, such as a 300-keV electron cryomicroscope (cryo-EM), the Titan Krios. Images are acquired on a direct-detection camera, the Falcon II, which has contributed to the cryo-EM revolution that is currently underway.
To answer questions about chromosome biology using cryo-ET, we have chosen budding and fission yeasts as our model organisms. Yeasts have numerous advantages, including their simplified cytology, simplified gene structure, and ease of growth and maintenance. The heroic efforts of our former PhD student Chen Chen have made cryo-ET of yeast routine. We have now exploited yeast to ask how cell-cycle, post-translational modifications, and architectural proteins influence chromosome structure.
Chromosomes are the first and final frontier of structural cell biology. They contain the instructions of life, which must be read, copied, and passed on to daughter cells. These activities must be done accurately by each of the trillions of cells in the human body. Mistakes can lead to many diseases, including cancers. How do cells access and use the information stored in chromosomes? How do they fold up the fundamental subunits called nucleosomes and then transport them into daughter cells? The answers to many of these fundamental questions will require knowledge of the organization, positions, and distributions of the subunits of chromosomes and their interaction partners, all within their native intracellular context.
The DNA and proteins that make up chromosomes are measured in nanometers, but they reside inside cells that are measured in micrometers. To map out the nanoscale organization of chromosomes inside their native cellular context, we use state-of-the-art electron tomography to generate 3-D images called “tomograms”. To minimize the fixation, dehydration, extraction, and staining artifacts associated with conventional electron microscopy (EM), we prepare and image our cells in a life-like frozen-hydrated state. This combination of 3-D nanoscale imaging and cryogenic sample handling is called cryo-electron tomography (cryo-ET). All our work has been done at the NUS Center for Bioimaging Sciences, which is equipped with top-of-the-line instruments, such as a 300-keV electron cryomicroscope (cryo-EM), the Titan Krios. Images are acquired on a direct-detection camera, the Falcon II, which has contributed to the cryo-EM revolution that is currently underway.
To answer questions about chromosome biology using cryo-ET, we have chosen budding and fission yeasts as our model organisms. Yeasts have numerous advantages, including their simplified cytology, simplified gene structure, and ease of growth and maintenance. The heroic efforts of our former PhD student Chen Chen have made cryo-ET of yeast routine. We have now exploited yeast to ask how cell-cycle, post-translational modifications, and architectural proteins influence chromosome structure.
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biorxiv(2024)
bioRxiv (Cold Spring Harbor Laboratory) (2024)
Corinne A Lutomski,Tarick J El-Baba,Carol V Robinson, Roland Riek,Sjors H W Scheres,Nieng Yan, Mohammed AlQuraishi,Lu Gan
Proceedings of the European Microscopy Congress 2020 (2021)
bioRxiv (Cold Spring Harbor Laboratory)pp.746982-51, (2021)
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