Session B43: Invited Session: Physical Mechanisms of Cell Growth

Processive motions of MreB micro-filaments coordinate cell wall growth

Rod-shaped bacteria elongate by the action of cell-wall synthesis complexes linked to underlying dynamic MreB filaments, but how these proteins function to allow continued elongation as a rod remains unknown. To understand how the movement of these filaments relates to cell wall synthesis, we characterized the dynamics of MreB and the cell wall elongation machinery using high-resolution particle tracking in \textit{Bacillus subtilis}. We found that both MreB and the elongation machinery move in linear paths across the cell, moving at similar rates ($\sim $20nm / second) and angles to the cell body, suggesting they function as single complexes. These proteins move circumferentially around the cell, principally perpendicular to its length. We find that the motions of these complexes are independent, as they can pause and reverse,and also as nearby complexes move independently in both directions across one surface of the cell. Inhibition of cell wall synthesis with antibiotics or depletions in the cell wall synthesis machinery blocked MreB movement, suggesting that the cell wall synthetic machinery is the motor in this system. We propose that bacteria elongate by the uncoordinated, circumferential movements of synthetic complexes that span the plasma membrane and insert radial hoops of new peptidoglycan during their transit.   

Spatial Patterning of Newly-Inserted Material during Bacterial Cell Growth

In the life cycle of a bacterium, rudimentary microscopy demonstrates that cell growth and elongation are essential characteristics of cellular reproduction. The peptidoglycan cell wall is the main load-bearing structure that determines both cell shape and overall size. However, simple imaging of cellular growth gives no indication of the spatial patterning nor mechanism by which material is being incorporated into the pre-existing cell wall. We employ a combination of high-resolution pulse-chase fluorescence microscopy, 3D computational microscopy, and detailed mechanistic simulations to explore how spatial patterning results in uniform growth and maintenance of cell shape. We show that growth is happening in discrete bursts randomly distributed over the cell surface, with a well-defined mean size and average rate. We further use these techniques to explore the effects of division and cell wall disrupting antibiotics, like cephalexin and A22, respectively, on the patterning of cell wall growth in \textit{E. coli.} Finally, we explore the spatial correlation between presence of the bacterial actin-like cytoskeletal protein, MreB, and local cell wall growth. Together these techniques form a powerful method for exploring the detailed dynamics and involvement of antibiotics and cell wall-associated proteins in bacterial cell growth.\\[4pt] In collaboration with Kerwyn Huang, Stanford University.   

Mechanics of cell division in fission yeast

Cytokinesis is the stage of cell division in which a cell divides into two. A paradigm of cytokinesis in animal cells is that the actomyosin contractile ring provides the primary force to squeeze the cell into two. In the fission yeast \textit{Schizosaccharomyces pombe}, cytokinesis also requires a actomyosin ring, which has been generally assumed to provide the force for cleavage. However, in contrast to animal cells, yeast cells assemble a cell wall septum concomitant with ring contraction and possess large (MPa) internal turgor pressure. Here, we show that the inward force generated by the division apparatus opposes turgor pressure; a decrease in effective turgor pressure leads to an increase in cleavage rate. We show that the ring cannot be the primary force generator. Scaling arguments indicate that the contractile ring can only provide a tiny fraction of the mechanical stress required to overcome turgor. Further, we show that cleavage can occur even in the absence of the contractile ring. Instead of the contractile ring, scaling arguments and modeling suggest that the large forces for cytokinesis are produced by the assembly of cell wall polymers in the growing septum.   

Decoding the topology of vascular organization

Distribution and structural networks permeate virtually all life, from the cellular to the organismic level. They have allowed organisms to grow in size and complexity by ensuring efficient distribution of nutrients and structural support. Given their importance, these vascular and structural webs have been under strong evolutionary selection and their form frequently reflects important aspects of their function. We discuss the design principles behind the evolution of the architecture and topology of vascular and structural networks and present some examples (leaf venation, arterial vasculature of the neocortex and others) that elucidate them.   

Mechanics and Dynamics of Plant Cell Division

The division of eukaryotic cells involves the assembly of complex cytoskeletal structures to exert the forces required for chromosome segregation and cytokinesis. In plants, tensional forces within the cytoskeleton constrain cells to divide according to a small number of area minimizing configurations. We have shown that the probability of observing a particular division configuration increases inversely with its relative area according to an exponential probability distribution known as the Gibbs measure. The distribution is universal up to experimental accuracy with a unique constant that applies for all plants studied irrespective of the shape and size of their cells. Using a maximum entropy formulation, we were able to demonstrate that the empirically observed division rule is predicted by the dynamics of the tense cytoskeletal elements controlling the positioning of the division plane. Finally, by framing this division rule as a dynamical system, we identified a broad class of attractors that are predictive of cell patterns observed in plants. Plant cell division thus offers a remarkable example of how interactions at the molecular level can lead to strikingly complex behaviors at the cellular and multicellular levels.