AbstractRegulation of the first committed step of peptidoglycan precursor synthesis by MurA‐enzyme homologs has recently taken center stage in many different bacteria. In different low‐GC Gram‐positive bacteria, regulation of this step has been shown to be regulated by phosphorylation of homologs of the IreB/ReoM regulatory protein by PASTA‐domain Ser/Thr‐protein kinases. In this issue, Mascari, Little, and Kristich determine this regulatory pathway and its links to resistance to cephalosporin β‐lactam antibiotics in the major human pathogen, Enterococcus faecalis (Efa). Unbiased genetic selections identified MurAA (MurA‐family homolog) as the downstream target of IreB regulation in the absence of the IreK Ser/Thr‐protein kinase. Physiological and biochemical approaches, including determination of MICs to ceftriaxone, Western blotting of MurAA cellular amounts, isotope incorporation into peptidoglycan sacculi, and thermal‐shift binding assays of purified proteins, demonstrated that unphosphorylated IreB, together with proteins MurAB (MurZ‐family homolog), and ReoY(Efa) negatively regulate MurAA stability and cellular amount by the ClpCP protease. Importantly, this paper supports the idea that ceftriaxone stimulates phosphorylation of IreB, which leads to increased cellular MurAA amount and precursor pathway flux required for E. faecalis cephalosporin resistance. Overall, findings in this paper significantly contribute to understanding variations of this central regulatory pathway in other low‐GC Gram‐positive bacteria.