title: Mechanical consequences of cell-wall turnover in the elongation of a Gram-positive bacterium.
abstract: A common feature of walled organisms is their exposure to osmotic forces that challenge the mechanical integrity of cells while driving elongation. Most bacteria rely on their cell wall to bear osmotic stress and determine cell shape. Wall thickness can vary greatly among species, with Gram-positive bacteria having a thicker wall than Gram-negative bacteria. How wall dimensions and mechanical properties are regulated and how they affect growth have not yet been elucidated. To investigate the regulation of wall thickness in the rod-shaped Gram-positive bacterium Bacillus subtilis, we analyzed exponentially growing cells in different media. Using transmission electron and epifluorescence microscopy, we found that wall thickness and strain were maintained even between media that yielded a threefold change in growth rate. To probe mechanisms of elongation, we developed a biophysical model of the Gram-positive wall that balances the mechanical effects of synthesis of new material and removal of old material through hydrolysis. Our results suggest that cells can vary their growth rate without changing wall thickness or strain by maintaining a constant ratio of synthesis and hydrolysis rates. Our model also indicates that steady growth requires wall turnover on the same timescale as elongation, which can be driven primarily by hydrolysis rather than insertion. This perspective of turnover-driven elongation provides mechanistic insight into previous experiments involving mutants whose growth rate was accelerated by the addition of lysozyme or autolysin. Our approach provides a general framework for deconstructing shape maintenance in cells with thick walls by integrating wall mechanics with the kinetics and regulation of synthesis and turnover.
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA, USA.
authors.forename: Gaurav
authors.initials: G
authors.lastname: Misra
authors.forename: Enrique R
authors.initials: ER
authors.lastname: Rojas
authors.forename: Ajay
authors.initials: A
authors.lastname: Gopinathan
authors.forename: Kerwyn Casey
authors.initials: KC
authors.lastname: Huang
mesh_terms: D001412:Bacillus subtilis / Q000166:cytology*; D001696:Biomechanical Phenomena; D049109:Cell Proliferation; D002473:Cell Wall / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D007700:Kinetics; D055595:Mechanical Phenomena; D059010:Single-Cell Analysis; D013314:Stress, Mechanical
chemical_list: D003470:Culture Media