Exploring molecular landscapes inside cells with in situ cryo-electron tomography
- Abstract number
- 354
- Corresponding Email
- [email protected]
- Session
- Stream 6 (Frontiers): Correlative Imaging of Organelle Organization and Architecture
- Authors
- Dr Benjamin Engel (1)
- Affiliations
-
1. Helmholtz Zentrum München
- Abstract text
Cells accomplish the biochemical reactions of life by concentrating their proteins into a variety of subcellular compartments called organelles. Our group explores the relationship between the form of the organelle and the function of its resident macromolecules. How does organelle architecture direct molecular function, and reciprocally, how do macromolecules sculpt and shape organelles? To investigate these questions, we use focused ion beam (FIB) milling of frozen cells followed by cryo-electron tomography to image macromolecules within their native cellular environment. Through a combination of nanometer-precision localization and high-resolution structural analysis, we aim to chart the molecular landscapes of organelles. Thanks to its superb cryo-EM contrast and textbook organelle architecture, the unicellular green alga Chlamydomonas is an ideal specimen for this approach. We have taken a holistic approach to survey the whole integrated “planimal”, with in situ molecular studies of the nuclear envelope, ER, Golgi, basal body apparatus (centrioles), and chloroplast. In this talk, I will provide an overview of some of these studies, touching on proteasome-rich degradation centers (Albert et al., 2017), the nuclear pore complex (Mosalaganti et al., 2018), COPI coats (Bykov et al., 2017), and the molecular organization of chloroplast’s thylakoid membranes and pyrenoid (Freeman Rosenzweig et al., 2017).
- References
Albert, S., Schaffer, M., Beck, F., Mosalaganti, S., Asano, S., Thomas, H.F., Plitzko, J.M., Beck, M., Baumeister, W., and Engel, B.D. Proteasomes tether to two distinct sites at the nuclear pore complex. Proc Natl Acad Sci U S A 114, 13726-13731 (2017).
Bykov, Y.S., Schaffer, M., Dodonova, S.O., Albert, S., Plitzko, J.M., Baumeister, W., Engel, B.D., and Briggs, J.A. The structure of the COPI coat determined within the cell. Elife 6, e32493 (2017).
Freeman Rosenzweig, E.S., Xu, B., Kuhn Cuellar, L., Martinez-Sanchez, A., Schaffer, M., Strauss, M., Cartwright, H.N., Ronceray, P., Plitzko, J.M., Forster, F., Wingreen, N.S., Engel, B.D., Mackinder, L.C.M., and Jonikas, M.C. The Eukaryotic CO2-Concentrating Organelle Is Liquid-like and Exhibits Dynamic Reorganization. Cell 171, 148-162.e119 (2017).
Mosalaganti, S., Kosinski, J., Albert, S., Schaffer, M., Strenkert, D., Salomé, P.A., Merchant, S.S., Plitzko, J.M., Baumeister, W., Engel, B.D., and Beck, M. In situ architecture of the algal nuclear pore complex. Nature Communications 9, 2361 (2018).