@article{TEXTUAL, recid = {10334}, author = {Ibrahim, Amany M. and Azam, Muhammad S. and Schneewind, Olaf and Missiakas, Dominique}, title = {Processing of LtaS restricts LTA assembly and YSIRK preprotein trafficking into &lt;i&gt;Staphylococcus aureus&lt;/i&gt; cross-walls}, journal = {mBio}, address = {2024-01-04}, number = {TEXTUAL}, abstract = {Septal membranes of Staphylococcus aureus serve as the site of secretion for precursors endowed with the YSIRK motif. Depletion of ltaS, a gene required for lipoteichoic acid (LTA) synthesis, results in the loss of restricted trafficking of YSIRK precursors to septal membranes. Here, we seek to understand the mechanism that ties LTA assembly and trafficking of YSIRK precursors. We confirm that catalytically inactive lipoteichoic acid synthase (LtaS)<sub>T300A</sub> does not support YSIRK precursor trafficking to septa. We hypothesize that the enzyme’s reactants [gentiobiosyldiacylglycerol (Glc<sub>2</sub>-DAG) and phosphatidylglycerol (PG)] or products [LTA and diacylglycerol (DAG)], not LtaS, must drive this process. Indeed, we observe that septal secretion of the staphylococcal protein A YSIRK precursor is lost in ypfP and ltaA mutants that produce glycerophosphate polymers [poly(Gro-P)] without the Glc<sub>2</sub>-DAG lipid anchor. These mutants display longer poly(Gro-P) chains, implying enhanced PG consumption and DAG production. Our experiments also reveal that in the absence of Glc<sub>2</sub>-DAG, the processing of LtaS to the extracellular catalytic domain, eLtaS, is impaired. Conversely, LTA polymerization is delayed in a strain producing LtaSS<sub>218P</sub>, a variant processed more slowly than LtaS. We conclude that Glc<sub>2</sub>-DAG binding to the enzyme couples catalysis by LtaS and the physical release of eLtaS. We propose a model for the temporal and localized assembly of LTA into cross-walls. When LtaS is not processed in a timely manner, eLtaS no longer diffuses upon daughter cell splitting, LTA assembly continues, and the unique septal-lipid pool, PG over DAG ratio, is not established. This results in profound physiological changes in S. aureus cells, including the inability to restrict the secretion of YSIRK precursors at septal membranes.}, url = {http://knowledge.uchicago.edu/record/10334}, }