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It consists of the actin\u2010like protein MreB, the PG synthase RodA\u2010PBP2 complex, as well as MreCD and RodZ. Although the loss or disruption of any elongasome component results in a loss of rod shape, previous studies found that a constitutively active RodA\u2010PBP2 complex can partially bypass the requirement of MreCD and RodZ for lateral PG synthesis and restore rod shape. However, how MreB is connected to RodA\u2010PBP2 under this situation and whether this linkage is important for elongasome activity in wild\u2010type cells remain unknown. Here, we report the isolation of additional RodA and PBP2 variants that can partially compensate for the absence of MreCD and RodZ in lateral PG synthesis. Taking advantage of these mutants and guided by an AlphaFold 3 structural model of the elongasome complex, we discover that both the cytoplasmic region of PBP2 and the C\u2010terminal tail of RodA interact with MreB. Moreover, disruption of these interactions results in a loss of rod shape, indicating that the interaction between MreB and RodA\u2010PBP2 is critical for elongasome function. Taken together, our results uncover how the MreB cytoskeleton is coupled to RodA\u2010PBP2 to organize lateral PG synthesis. These findings provide mechanistic insights into cell wall biogenesis in bacteria and offer strategies for the development of new antibiotics targeting the elongasome.<\/jats:p>","DOI":"10.1002\/mlf2.70079","type":"journal-article","created":{"date-parts":[[2026,5,18]],"date-time":"2026-05-18T11:33:04Z","timestamp":1779103984000},"page":"325-338","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Direct interaction between MreB and the RodA\u2010PBP2 complex organizes lateral cell wall synthesis in\n                    <i>Escherichia coli<\/i>"],"prefix":"10.1002","volume":"5","author":[{"given":"Rui","family":"Zhan","sequence":"first","affiliation":[{"name":"State Key Laboratory of Metabolism and Regulation in Complex Organisms College of Life Sciences, Wuhan University Wuhan China"},{"name":"Hubei Key Laboratory of Cell Homeostasis College of Life Sciences, Wuhan University Wuhan 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