Deep single-cell phenotyping to identify governing principles and mechanisms of the subcellular organization of bacterial replication

Introduction

Modern metagenomics has opened our eyes to the immense bacterial diversity that exists both among and within us. Despite this diversity, all bacteria share the basic challenge of organizing the various processes that ensure their faithful replication.

Bacterial Processes

All bacterial cells need to:

1. Metabolize nutrients
2. Generate building blocks
3. Maintain their shape and size
4. Replicate and segregate their chromosomes
5. Synthesize cell walls and membranes
6. Divide to give rise to daughter cells

At present, we do not understand how bacteria integrate all these processes in their small cellular compartments.

Intriguing Simplicity

What makes this question even more intriguing is that bacteria represent simple forms of proliferating cells, without additional layers of internal organization (e.g., membrane-enclosed organelles) or cell cycle regulation (e.g., cyclins and cyclin-dependent kinases) seen in eukaryotic cells.

Research Goal

My goal is to address this gap by uncovering the internal architecture of bacterial replication and identifying the molecular mechanisms that underlie it.

Methodology

I will use a high-throughput single-cell phenomics approach that I developed, which provides high-content, quantitative cell biological information. By applying this approach across different levels of bacterial diversity (both within and across species, beyond the small number of currently existing model species), I aim to identify general and species-specific principles for the subcellular organization of replication in bacteria.

Expected Outcomes

This analysis will also enable the identification of key factors involved in establishing these governing principles, which will be functionally characterized further to provide a unique overview of the molecular mechanisms that determine the spatial organization of bacterial replication.

If successful, this project will transform our understanding of bacterial cell biology by expanding it beyond current textbook standards and providing us with the blueprints and design principles of bacterial cells.