| Texto completo | |
| Autor(es): |
Shirakawaa, Karina T.
;
Salaa, Fernanda Angelica
;
Miyachiroc, Mayara M.
;
Jobc, Viviana
;
Trindadea, Daniel Maragno
;
Dessena, Andrea
;
Mayer, Christoph
;
Karls, Eberhard
Número total de Autores: 8
|
| 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 |