The MurG glycosyltransferase provides an oligomeric scaffold for the cytoplasmic steps of peptidoglycan biosynthesis in the human pathogen Bordetella pertussis

Laddomada, Federica; Miyachiro, Mayara M.; Jessop, Matthew; Patin, Delphine; Job, Viviana; Mengin‐Lecreulx, Dominique; Roy, Aline Le; Ebel, Christine; Breyton, Cécile; Gutsche, Irina; Dessen, Andréa

doi:10.1038/s41598-019-40966-z

Cited by 16 publications

(18 citation statements)

References 55 publications

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“…4A). MurG has been shown to act as a scaffold for PG biosynthesis in Bordetella pertussis and Thermotoga maritima (64,65) and could provide a similar function in C. crescentus, though it remains unclear if the changes we observe for MurG are due to absence of a direct interaction with GroESL, or perhaps loss of an upstream signal required for PG biosynthetic subcomplex assembly. Depletion of PG precursors, which may occur through a reduction in pathway protein function, results in filamentation to conserve limited resources and prevent overinvestment in the intensive process of building new cell poles (51).…”

Section: Discussionmentioning

confidence: 89%

The Chaperonin GroESL Facilitates Caulobacter crescentus Cell Division by Supporting the Functions of the Z-Ring Regulators FtsA and FzlA

Schroeder

Heinrich

Neuwirth

et al. 2021

mBio

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The highly conserved chaperonin GroESL performs a crucial role in protein folding; however, the essential cellular pathways that rely on this chaperone are underexplored. Loss of GroESL leads to severe septation defects in diverse bacteria, suggesting the folding function of GroESL may be integrated with the bacterial cell cycle at the point of cell division. Here, we describe new connections between GroESL and the bacterial cell cycle using the model organism Caulobacter crescentus. Using a proteomics approach, we identify candidate GroESL client proteins that become insoluble or are degraded specifically when GroESL folding is insufficient, revealing several essential proteins that participate in cell division and peptidoglycan biosynthesis. We demonstrate that other cell cycle events, such as DNA replication and chromosome segregation, are able to continue when GroESL folding is insufficient. We further find that deficiency of two FtsZ-interacting proteins, the bacterial actin homologue FtsA and the constriction regulator FzlA, mediate the GroESL-dependent block in cell division. Our data show that sufficient GroESL is required to maintain normal dynamics of the FtsZ scaffold and divisome functionality in C. crescentus. In addition to supporting divisome function, we show that GroESL is required to maintain the flow of peptidoglycan precursors into the growing cell wall. Linking a chaperone to cell division may be a conserved way to coordinate environmental and internal cues that signal when it is safe to divide. IMPORTANCE All organisms depend on mechanisms that protect proteins from misfolding and aggregation. GroESL is a highly conserved molecular chaperone that functions to prevent protein aggregation in organisms ranging from bacteria to humans. Despite detailed biochemical understanding of GroESL function, the in vivo pathways that strictly depend on this chaperone remain poorly defined in most species. This study provides new insights into how GroESL is linked to the bacterial cell division machinery, a crucial target of current and future antimicrobial agents. We identify a functional interaction between GroESL and the cell division proteins FzlA and FtsA, which modulate Z-ring function. FtsA is a conserved bacterial actin homologue, suggesting that as in eukaryotes, some bacteria exhibit a connection between cytoskeletal actin proteins and chaperonins. Our work further defines how GroESL is integrated with cell wall synthesis and illustrates how highly conserved folding machines ensure the functioning of fundamental cellular processes during stress.

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Section: Discussionmentioning

confidence: 89%

The Chaperonin GroESL Facilitates Caulobacter crescentus Cell Division by Supporting the Functions of the Z-Ring Regulators FtsA and FzlA

Schroeder

Heinrich

Neuwirth

et al. 2021

mBio

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“…Although structural studies have provided no direct evidence, a highly oligomeric state of MurG has consistently been proposed as a functional unit by biochemical, biophysical and cellular studies (Laddomada et al, 2019;Ha et al, 1999).…”

Section: Abmurg Forms Hexameric Homo-oligomeric Complex Structuresmentioning

confidence: 99%

“…Owing to the critical activity of MurG in bacterial cell-wall synthesis and its attractiveness as an antibiotic target, structural and biochemical studies have been performed in order to understand its molecular mechanism (Ha et al, 1999;Mohammadi et al, 2007;van den Brink-van der Laan et al, 2003;Hu et al, 2004). Although previous studies of Escherichia coli MurG (ecMurG) have shown how MurG functions in the membrane and how its active site recognizes substrates (Ha et al, 2000;Hu et al, 2003), the molecular mechanism of the oligomeric scaffolding assembly of MurG, as identified in several bacterial species in recent studies (Laddomada et al, 2019), has remained unclear. This study characterized MurG from A. baumannii (abMurG).…”

Section: Introductionmentioning

confidence: 99%

Putative hexameric glycosyltransferase functional unit revealed by the crystal structure of Acinetobacter baumannii MurG

Jung

Kwon

Kim

et al. 2021

Int Union Crystallogr J

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Lipid II, the main component of the bacterial cell wall, is synthesized by the addition of UDP-N-acetylglucosamine to the UDP-N-acetylmuramic acid pentapeptide catalyzed by the glycosyltransferase MurG. Owing to its critical role in cell-wall biosynthesis, MurG is considered to be an attractive target for antibacterial agents. Although the Mur family ligases have been extensively studied, the molecular mechanism of the oligomeric scaffolding assembly of MurG remains unclear. In this study, MurG from Acinetobacter baumannii (abMurG), a human pathogen, was characterized and its hexameric crystal structure was unveiled; this is the first homo-oligomeric structure to be described in the MurG family and the Mur family. Homogeneous protein samples were produced for structural studies using size-exclusion chromatography, the absolute molecular mass was calculated via multi-angle light scattering, and protein–protein interactions were analyzed using the PDBePISA server. abMurG was found to form homo-oligomeric complexes in solution, which might serve as functional units for the scaffolding activity of MurG. Furthermore, analysis of this structure revealed the molecular assembly mechanism of MurG. This structural and biochemical study elucidated the homo-oligomerization mechanism of MurG and suggests a new potential antibiotic target on MurG.

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“…Our data suggests that MurG solubility and localization may also respond to GroESL-mediated folding ( Figure 4A). MurG has been shown to act as a scaffold for PG biosynthesis in Bordetella pertussis and Thermotoga maritima (63,64) processes are protected during high unfolding demand. Discerning these interactions will be important to understanding how organisms balance growth and division when surviving stress.…”

Section: Crescentus and Has Identified Several New Interactions Betwmentioning

confidence: 99%

The chaperonin GroESL facilitatesCaulobacter crescentuscell division by supporting the function of the actin homologue FtsA

Schroeder

Heinrich

Neuwirth

et al. 2020

Preprint

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The highly conserved chaperonin GroESL performs a crucial role in protein folding, however the essential cellular pathways that rely on this chaperone are underexplored. Loss of GroESL leads to severe septation defects in diverse bacteria, suggesting the folding function of GroESL may be integrated with the bacterial cell cycle at the point of cell division. Here, we describe new connections between GroESL and the bacterial cell cycle, using the model organism Caulobacter crescentus. Using a proteomics approach, we identify candidate GroESL client proteins that become insoluble or are degraded specifically when GroESL folding is insufficient, revealing several essential proteins that participate in cell division and peptidoglycan biosynthesis. We demonstrate that other cell cycle events such as DNA replication and chromosome segregation are able to continue when GroESL folding is insufficient, and find that deficiency of the bacterial actin homologue FtsA function mediates the GroESL-dependent block in cell division. Our data suggest that a GroESL-FtsA interaction is required to maintain normal dynamics of the FtsZ scaffold and divisome functionality in C. crescentus. In addition to supporting FtsA function, we show that GroESL is required to maintain the flow of peptidoglycan precursors into the growing cell wall. Linking a chaperone to cell division may be a conserved way to coordinate environmental and internal cues that signal when it is safe to divide.ImportanceAll organisms depend on mechanisms that protect proteins from misfolding and aggregation. GroESL is a highly conserved molecular chaperone that functions to prevent protein aggregation in organisms ranging from bacteria to humans. Despite detailed biochemical understanding of GroESL function, the in vivo pathways that strictly depend on this chaperone remain poorly defined in most species. This study provides new insights into how GroESL is linked to the bacterial cell division machinery, a crucial target of current and future antimicrobial agents. We identify a functional interaction between GroESL and FtsA, a conserved bacterial actin homologue, suggesting that as in eukaryotes, some bacteria exhibit a connection between cytoskeletal actin proteins and chaperonins. Our work further defines how GroESL is integrated with cell wall synthesis, and illustrates how highly conserved folding machines ensure the functioning of fundamental cellular processes during stress.

show abstract

The MurG glycosyltransferase provides an oligomeric scaffold for the cytoplasmic steps of peptidoglycan biosynthesis in the human pathogen Bordetella pertussis

Cited by 16 publications

References 55 publications

The Chaperonin GroESL Facilitates Caulobacter crescentus Cell Division by Supporting the Functions of the Z-Ring Regulators FtsA and FzlA

The Chaperonin GroESL Facilitates Caulobacter crescentus Cell Division by Supporting the Functions of the Z-Ring Regulators FtsA and FzlA

Putative hexameric glycosyltransferase functional unit revealed by the crystal structure of Acinetobacter baumannii MurG

The chaperonin GroESL facilitatesCaulobacter crescentuscell division by supporting the function of the actin homologue FtsA

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