The cell cycle in Staphylococcus aureus is regulated by an amidase that controls peptidoglycan synthesis

Abstract

13Bacteria are protected by a polymer of peptidoglycan that serves as an exoskeleton. In 14 Staphylococcus aureus, the enzymes that assemble peptidoglycan move during the cell cycle from 15 the periphery, where they are active during growth, to the division site where they build the 16 partition between daughter cells. But how peptidoglycan synthesis is regulated throughout the cell 17 cycle is not understood. Here we identify a membrane protein complex that spatially regulates S. 18 aureus peptidoglycan synthesis. This complex consists of an amidase that removes peptide chains 19 from uncrosslinked peptidoglycan and a partner protein that controls its activity. Typical amidases 20 act after cell division to hydrolyze peptidoglycan between daughter cells so they can separate. 21 130 peptidoglycan that is not, or at least not extensively, crosslinked. 131Slowing peptidoglycan synthesis rescues lytH deletion defects 132 We next investigated how LytH could regulate cell division. The division defects resulting 133 from lytH deletion implied LytH is important for FtsZ assembly. We found using a functional 134 FtsZ-sGFP fusion 5 that FtsZ is frequently mislocalized in ∆lytH cells ( Fig. 4a and Supplementary 135 157 misplaced septa or peripheral puncta ( Fig. 5a and Supplementary Fig. 14). By measuring the ratio 158

Structure and reconstitution of a hydrolase complex that may release peptidoglycan from the membrane after polymerization

“…The final deletions were confirmed by colony PCR and sequencing. Phage transductions were performed using a previously published protocol 39 .…”

The Sle1 Cell Wall Amidase Is Essential for β-Lactam Resistance in Community-Acquired Methicillin-Resistant Staphylococcus aureus USA300

“…Consistent with this finding, Matias and Beveridge (24) proposed that cell wall autolysins are required for synchronized growth of the septum. Intriguingly, a recent report suggests that another S. aureus cell wall amidase, LytH, is involved in controlling the spatial distribution of peptidoglycan synthases to ensure that cell expansion is coordinated with cell division (37). Hence, the activities of cell wall hydrolases and PBPs may be tightly linked to balance peptidoglycan synthesis with autolytic degradation, and the binding of ␤-lactam antibiotics PBPs may perturb this delicate balance.…”

Deletion of the novel gene mother cell lysis X results in Cry1Ac encapsulation in the Bacillus thuringiensis HD73

“…The endopeptidase EagA is regulated through bacterial second messenger molecule c-di-GMP signaling in Erwinia amylovora (Kharadi and Sundin, 2020 ). Protein interactions with hydrolases can activate (Uehara et al, 2009(Uehara et al, , 2014Yang et al, 2011;Domínguez-Cuevas et al, 2013;Meisner et al, 2013;Do et al, 2020b) or inhibit hydrolase activities (Clarke et al, 2010). The mother cell of the HD ( mclX) strain did not lyse because the cwlC expression decreased (Figure 4A).…”