Outer membrane lipoprotein NlpI scaffolds peptidoglycan hydrolases within multi‐enzyme complexes in Escherichia coli

• Manuel Banzhaf ^[4] ; Hamish CL Yau ^[1] ; Jolanda Verheul ^[2] ; Adam Lodge ^[1] ; George Kritikos ^[5] ; André Mateus ^[5] ; Baptiste Cordier ^[6] ; Ann Kristin Hov ^[7] ; Frank Stein ^[5] ; Morgane Wartel ^[5] ; Manuel Pazos ^[1] ; Alexandra S Solovyova ^[8] ; Eefjan Breukink ^[3] ; Sven van Teeffelen ^[6] ; Mikhail M Savitski ^[9] ; Tanneke den Blaauwen ^[2] ; Athanasios Typas ^[9] ; Waldemar Vollmer ^[1]
  1. [1] Newcastle University

     Newcastle University

     Reino Unido

     
  2. [2] University of Amsterdam

     University of Amsterdam

     Países Bajos

     
  3. [3] Utrecht University

     Utrecht University

     Países Bajos

     
  4. [4] 1 European Molecular Biology Laboratory Genome Biology Unit Heidelberg Germany; 8Present address: Institute of Microbiology & Infection and School of Biosciences University of Birmingham Edgbaston Birmingham UK
  5. [5] 1 European Molecular Biology Laboratory Genome Biology Unit Heidelberg Germany
  6. [6] 4 Microbial Morphogenesis and Growth Lab Institut Pasteur Paris France
  7. [7] 1 European Molecular Biology Laboratory Genome Biology Unit Heidelberg Germany; 11Present address: École polytechnique fédérale de Lausanne SV IBI‐SV UPDALPE, AAB 013 Lausanne Switzerland
  8. [8] 5 Newcastle University Protein and Proteome Analysis Newcastle Upon Tyne UK
  9. [9] 1 European Molecular Biology Laboratory Genome Biology Unit Heidelberg Germany; 7 European Molecular Biology Laboratory Structural & Computational Unit Heidelberg Germany

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• Localización: EMBO journal: European Molecular Biology Organization, ISSN 0261-4189, Vol. 39, Nº. 5, 2020
• Idioma: inglés
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  • The peptidoglycan (PG) sacculus provides bacteria with the mechanical strength to maintain cell shape and resist osmotic stress. Enlargement of the mesh‐like sacculus requires the combined activity of peptidoglycan synthases and hydrolases. In Escherichia coli, the activity of two PG synthases is driven by lipoproteins anchored in the outer membrane (OM). However, the regulation of PG hydrolases is less well understood, with only regulators for PG amidases having been described. Here, we identify the OM lipoprotein NlpI as a general adaptor protein for PG hydrolases. NlpI binds to different classes of hydrolases and can specifically form complexes with various PG endopeptidases. In addition, NlpI seems to contribute both to PG elongation and division biosynthetic complexes based on its localization and genetic interactions. Consistent with such a role, we reconstitute PG multi‐enzyme complexes containing NlpI, the PG synthesis regulator LpoA, its cognate bifunctional synthase, PBP1A, and different endopeptidases. Our results indicate that peptidoglycan regulators and adaptors are part of PG biosynthetic multi‐enzyme complexes, regulating and potentially coordinating the spatiotemporal action of PG synthases and hydrolases.