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

Autor(es)

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

• Tipo de documento: Artigo Científico
• Fonte: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA; v. 120, n. 21, p. 10-pg., 2023-05-15.

Resumo

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)

• Processo FAPESP: 11/52067-6 - Estruturação de complexos macromoleculares da parede bacteriana: biossíntese e virulência
• Beneficiário: Andrea Dessen de Souza e Silva
• Modalidade de apoio: Auxílio à Pesquisa - Programa SPEC
• Processo FAPESP: 17/12436-9 - ANTIBIO-BAC: a parede bacteriana como alvo para o desenvolvimento de novos agentes antimicrobianos
• Beneficiário: Andrea Dessen de Souza e Silva
• Modalidade de apoio: Auxílio à Pesquisa - Programa SPEC
• Processo FAPESP: 18/16346-7 - Estudo das interações diretas, cristalização e estruturas dos complexos binários entre as enzimas Mur de Streptococcus pneumoniae
• Beneficiário: Karina Tamie Shirakawa
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 20/01286-9 - Estudo bioquímico, biofísico e estrutural de um complexo de enzimas Mur do operon de Bordetella pertussis
• Beneficiário: Karina Tamie Shirakawa
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto