General principles for the formation and proliferation of a wall-free (L-form) state in bacteria

The peptidoglycan cell wall is a defining structural feature of the bacterial-kingdom. Curiously, some bacteria have the ability to switch to a wall-free or L-form state. Although known for decades, the general properties of L-forms are poorly understood, largely due to the lack of their systematic analysis in the molecular biology era. Here we show that inhibition of the peptidoglycan precursor synthesis promotes the generation of L-forms from both Gram-positive and Gram-negative bacteria. We show that L-forms generated have in common a mechanism of proliferation involving membrane blebbing and tubulation, which is dependant on an altered rate of membrane synthesis. Crucially, this mode of proliferation is independent of the essential FtsZ-based division machinery. Our results suggest that the L-form mode of proliferation is conserved across the bacterial-kingdom, reinforcing the idea that it could have been used in primitive cells, and opening up its use in the generation of synthetic cells.