Architecture and genomic arrangement of the MurE-MurF bacterial cell wall biosynthesis complex

Author(s)

Shirakawaa, Karina T. ; Salaa, Fernanda Angelica ; Miyachiroc, Mayara M. ; Jobc, Viviana ; Trindadea, Daniel Maragno ; Dessena, Andrea ; Mayer, Christoph ; Karls, Eberhard

• Document type: Journal article
• Source: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA; v. 120, n. 21, p. 10-pg., 2023-05-15.

Abstract

We acknowledge support to A.D. from the Laboratoire International Associe BACWALL(CNRS), and grants from the Sao Paulo Research Foundation (FAPESP) 2011/52067-6 and 2017/12436-9. K.T.S. was supported by fellowships 2018/16346-7 and 2020/01286-9 from FAPESP, the CNPq, and CAPES.This work made use of the MANACa beamline of the Brazilian Synchrotron Peptidoglycan (PG) is a central component of the bacterial cell wall, and the disruption of its biosynthetic pathway has been a successful antibacterial strategy for decades. PG biosynthesis is initiated in the cytoplasm through sequential reactions catalyzed by Mur enzymes that have been suggested to associate into a multimembered complex. This idea is supported by the observation that in many eubacteria, mur genes are present in a single operon within the well conserved dcw cluster, and in some cases, pairs of mur genes are fused to encode a single, chimeric polypeptide. We performed a vast genomic analysis using >140 bacterial genomes and mapped Mur chimeras in numerous phyla, with Proteobacteria carrying the highest number. MurE-MurF, the most prevalent chimera, exists in forms that are either directly associated or separated by a linker. The crystal structure of the MurE-MurF chimera from Bordetella pertussis reveals a head-to-tail, elongated architecture supported by an interconnecting hydrophobic patch that stabilizes the positions of the two proteins. Fluorescence polarization assays reveal that MurE-MurF interacts with other Mur ligases via its central domains with KDs in the high nanomolar range, backing the existence of a Mur complex in the cytoplasm. These data support the idea of stronger evolutionary constraints on gene order when encoded proteins are intended for association, establish a link between Mur ligase interaction, complex assembly and genome evolution, and shed light on regulatory mechanisms of protein expression and stability in pathways of critical importance for bacterial survival. (AU)

• FAPESP's process: 11/52067-6 - Assembly and structure of macromolecular complexes involved in bacterial cell wall: biosynthesis and virulence
• Grantee: Andrea Dessen de Souza e Silva
• Support Opportunities: Research Projects - SPEC Program
• FAPESP's process: 17/12436-9 - ANTIBIO-BAC: exploring the bacterial cell wall as a target for novel antibiotherapies
• Grantee: Andrea Dessen de Souza e Silva
• Support Opportunities: Research Projects - SPEC Program
• FAPESP's process: 18/16346-7 - Study of the direct interactions, crystallization and structure of the binary complexes between Mur enzymes from Streptococcus pneumoniae
• Grantee: Karina Tamie Shirakawa
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 20/01286-9 - Biochemical, biophysical and structural study of a Mur enzyme complex from the operon of Bordetella pertussis
• Grantee: Karina Tamie Shirakawa
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)