Our research program fits within one of the grand challenges of synthetic biology: integrating individual molecular systems into functional modules towards building a synthetic cell from the bottom-up. The achievement of this goal requires, among other things, a profound mechanistic understanding of how the elements of essential cellular machines are organized, in time and space, and coordinated one to another, as a network of multiple interactions to function in the crowded and phase-separated cell interior.
We have selected the bacterial division machinery – the divisome – as the system to address these fundamental questions. Cytokinesis in bacteria is known in sufficient detail to provide a comprehensive list of its molecular effectors and their biochemical properties; therefore, it seems reasonable to expect that the application of bottom-up synthetic strategy to construct minimal divisomes in the test tube is feasible and will yield functional assemblies. This synthetic approach will help us support conclusions already derived from cellular and molecular analysis and, therefore, complete our understanding of how the bacterial division works.
Keywords: cellular biochemistry, molecular biophysics, bottom-up synthetic biology, protein science, molecular interactions, macromolecular crowding, macromolecular phase separation, biomolecular condensates, biological self-organization, cell division, bacteria
- Molecular interactions in bacterial division: We aim at obtaining a complete biochemical description of how the FtsZ protein – the central element of the divisome in most bacteria – and the negative and positive regulators of division ring stability work together as an integrated system of molecular interactions, including those involving the lipid membrane and the bacterial chromosome. This knowledge will shed light on the molecular mechanisms responsible for coordinating cell cycle events in bacteria.
- Reconstructing bacterial division in cytomimetic environments: We study the activities, interactions, and assembly properties of FtsZ in minimal membrane systems and artificial cell systems, to define more precise conditions to reconstruct a growing number of divisome subsets capable of performing cell division functions in controlled cell-like environments, in the absence of cells. The final aim is to integrate these active divisome modules in artificial cells to lead to their autonomous division eventually.
- Macromolecular crowding and phase separation in bacterial division: We aim to define how physicochemical elements of intracellular complexity (macromolecular crowding, surface interactions, and – especially – biomolecular condensation mediated by phase separation) affect the biochemical reactivity (protein-protein, protein-DNA, and protein-DNA-membrane interactions) and spatiotemporal organization underlying the operation of minimal divisome machines.
- Enabling tools: We apply and develop front-line protein biochemistry, molecular interactions, and membrane reconstitution tools, combined with cutting-edge microsystems, and fluorescence micro-spectroscopy technologies, and facsimile cell conditions.
- Exploitation in antimicrobial discovery and technology: The knowledge and technologies acquired are being used to explore the design of novel assays to curb bacterial proliferation and the production of protein materials and devices with technological added value.
- European Synthetic Cell Initiative and CSyCell: Our laboratory coordinates the Spanish node of the European Synthetic Cell initiative, a multidisciplinary effort to build a functioning synthetic cell from the bottom-up. Achievement this challenge will significantly contribute to understanding how cells work, define life's basic principles, and lead to synthetic-cell technologies to solve health and environmental problems. In this line, we coordinate the efforts to assemble the CSIC minimal synthetic life program - CSyCell - which is one of the potential CSIC challenges for 2020-2040.
- BIOINTERACT-CIB: Our laboratory acts as the scientific coordinator of the Molecular Interactions Facility at the CIB Margarita Salas, a national reference laboratory in the study of biomolecular interactions with physiological, biotechnological, or biomedical relevance, using analytical ultracentrifugation and complementary tools (light scattering, fluorescence spectroscopy, and optical biosensing, among others).