InstructThe latest webinar in the Instruct-ERIC Structure Meets Function series includes speakers from Instruct-CZ. The webinar series offers an insight into the very cutting edge of structural biology research, utilising the latest techniques available through Instruct-ERIC facilities and centres.
This month we have two speakers, providing their experience and expertise with the services and technologies available at Instruct-CZ. Jiri Novacek of CEITEC introduces the solutions and services available for remote access of machinery at the facility, and data accessibility. Elda Bauda then provides insight into their Instruct internship at CEITEC, utilising Cryo-ET to characterise the structure of sporulating bacterial cells.
Speaker: Jiri Novacek - CEITEC
Title: SIP - platform for management of the scientific data life-cycle and its implementation in mid-sized cryo-EM service laboratory
Abstract: We present a comprehensive workflow that facilitates the management of raw scientific data. Our approach has been tested using electron cryo-microscopy (cryo-EM), which, like other imaging methods, generates substantial amounts of raw data per dataset. The workflow is built on the iRODS federated cloud system and incorporates previously developed tools for data publication. These components are orchestrated by a Python-based system to automate the indexing of data to remote cloud storage and, alternatively, its archival for long-term preservation. Additionally, the workflow includes the deposition of metadata into a public database, with release following the expiration of an embargo period. Furthermore, we have implemented real-time data analysis of single-particle cryo-EM data at a remote high-performance computing (HPC) center, eliminating the need for computational resources within the service laboratory.
Speaker: Elda Bauda - Université de Genève
Title: Ultrastructural assembly of protective bacterial spore structures revealed by cryo-ET
Abstract: Bacterial spores are dormant cells that are highly resistant to antibiotics, disinfectants, radiation, and high temperatures. This resilience, which can pose risks to public health and food safety, arises from molecular assemblies that protect the cell and its genetic material. These include the compaction of the DNA in the spore core and a multi-protein shell known as the coat.
To investigate the formation of these protective structures, we have implemented in situ cryo-electron tomography (cryo-ET) on Bacillus subtilis lamellae generated by cryo-focused ion beam milling (cryo-FIBM).
Our findings reveal that, at early stages of spore development, the chromosome adopts a toroidal organization, while coat proteins assemble at the spore surface into complex stacks of both amorphous and structured layers with varying dimensions and compositions.
Further analysis identified distinct regions within the nascent coat, involving key morphogenetic proteins, laying the groundwork for unraveling the molecular mechanisms behind bacterial spore resistance.
