Peptidoglycan maturation controls spatiotemporal organisation of outer membrane proteins in Escherichia coli

Linkages between the outer membrane of Gram-negative bacteria and the peptidoglycan layer are crucial to the maintenance of cellular integrity and enable survival in challenging environments[1][1]–[5][2]. The functionality of the outer membrane relies on outer membrane proteins (OMPs), which are inserted by the β-barrel assembly machine, BAM[6][3], [7][4]. Previous work has shown that growing Escherichia coli cells segregate old OMPs towards the poles by an unknown mechanism[8][5]. Here, we demonstrate that peptidoglycan underpins the spatiotemporal organisation of OMPs. Mature, tetrapeptide-rich peptidoglycan binds to BAM components and suppresses OMP foldase activity. Nascent peptidoglycan, which is enriched in pentapeptides and concentrated at septa[9][6], associates with BAM poorly and has little impact on its activity, leading to preferential insertion of OMPs at division sites. Synchronising OMP biogenesis to cell wall growth enables bacteria to replenish their OMPs by binary partitioning. Our study reveals that Gram-negative bacteria coordinate the assembly of two major cell envelope layers by rendering OMP biogenesis responsive to peptidoglycan maturation. This coordination offers new possibilities for the design of antibiotics that disrupt cell envelope integrity. ### Competing Interest Statement Dawei Sun is an employee of Genentech, a member of the Roche Group, and a shareholder in Roche. [1]: #ref-1 [2]: #ref-5 [3]: #ref-6 [4]: #ref-7 [5]: #ref-8 [6]: #ref-9