Cryo-electron tomography of bacterial minicells

Abstract

Pas de résuméThe following thesis recounts work in which cryo-electron tomography and subtomogramaveraging were applied to the study of chemosensory arrays in E. coli minicells. The focus ofthe work is the optimal application of subtomogram averaging to this biological system. Inparticular, the thesis draws upon recent developments in subtomogram averagingmethodology to overcome challenges associated with image processing in cryo-electrontomography. In the introductory chapter the use of cryo-electron tomography for in situstructural biology is introduced. Work describing chemosensory core-signalling unit structurewhich also introduces the biology of the system is presented in Chapter 2. A brief foray intothe description of an observed alternative chemosensory array ultrastructure constitutesChapter 3. Image analysis methodology in structural cryo-electron tomography has continuedto develop rapidly from 2017-2020, the period during which this work was undertaken. Assuch, Chapter 4 describes technical contributions which facilitated the application of state-of-the-art subtomogram averaging methodology to the study of chemosensory arrays and otherchallenging structural targets. Finally, Chapter 5 introduces promising first results on datacollected in the final stages of this thesis in the context of avenues for further study anddiscusses the future of cryo-electron tomography