Hydrolysis of peptidoglycan is modulated by amidation of meso-diaminopimelic acid and Mg(2+) in Bacillus subtilis

Alex Dajkovic, Benoit Tesson, Smita Chauhan, Pascal Courtin, Ruth Keary, Pierre Flores, Christian Marlière, Sérgio R Filipe, Marie-Pierre Chapot-Chartier, Rut Carballido-Lopez

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The ability of excess Mg(2+) to compensate the absence of cell wall related genes in Bacillus subtilis has been known for a long time, but the mechanism has remained obscure. Here, we show that the rigidity of wild-type cells remains unaffected with excess Mg(2+) , but the proportion of amidated meso-diaminopimelic (mDAP) acid in their peptidoglycan (PG) is significantly reduced. We identify the amidotransferase AsnB as responsible for mDAP amidation and show that the gene encoding it is essential without added Mg(2+) . Growth without excess Mg(2+) causes ΔasnB mutant cells to deform and ultimately lyse. In cell regions with deformations, PG insertion is orderly and indistinguishable from the wild-type. However, PG degradation is unevenly distributed along the sidewalls. Furthermore, ΔasnB mutant cells exhibit increased sensitivity to antibiotics targeting the cell wall. These results suggest that absence of amidated mDAP causes a lethal deregulation of PG hydrolysis that can be inhibited by increased levels of Mg(2+) . Consistently, we find that Mg(2+) inhibits autolysis of wild-type cells. We suggest that Mg(2+) helps to maintain the balance between PG synthesis and hydrolysis in cell wall mutants where this balance is perturbed in favor of increased degradation.

Original languageEnglish
Pages (from-to)972-988
Number of pages17
JournalMolecular Microbiology
Volume104
Issue number6
DOIs
Publication statusPublished - 24 Apr 2017

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Diaminopimelic Acid
Peptidoglycan
Bacillus subtilis
Hydrolysis
Cell Wall
Autolysis
Genes
Anti-Bacterial Agents
Growth

Keywords

  • GRAM-POSITIVE BACTERIA
  • WALL TEICHOIC-ACIDS
  • SOLID-STATE NMR
  • D-AMINO ACIDS
  • CELL-WALL
  • ESCHERICHIA-COLI
  • AUTOLYTIC SYSTEM

Cite this

Dajkovic, A., Tesson, B., Chauhan, S., Courtin, P., Keary, R., Flores, P., ... Carballido-Lopez, R. (2017). Hydrolysis of peptidoglycan is modulated by amidation of meso-diaminopimelic acid and Mg(2+) in Bacillus subtilis. Molecular Microbiology, 104(6), 972-988. https://doi.org/10.1111/mmi.13673
Dajkovic, Alex ; Tesson, Benoit ; Chauhan, Smita ; Courtin, Pascal ; Keary, Ruth ; Flores, Pierre ; Marlière, Christian ; Filipe, Sérgio R ; Chapot-Chartier, Marie-Pierre ; Carballido-Lopez, Rut. / Hydrolysis of peptidoglycan is modulated by amidation of meso-diaminopimelic acid and Mg(2+) in Bacillus subtilis. In: Molecular Microbiology. 2017 ; Vol. 104, No. 6. pp. 972-988.
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abstract = "The ability of excess Mg(2+) to compensate the absence of cell wall related genes in Bacillus subtilis has been known for a long time, but the mechanism has remained obscure. Here, we show that the rigidity of wild-type cells remains unaffected with excess Mg(2+) , but the proportion of amidated meso-diaminopimelic (mDAP) acid in their peptidoglycan (PG) is significantly reduced. We identify the amidotransferase AsnB as responsible for mDAP amidation and show that the gene encoding it is essential without added Mg(2+) . Growth without excess Mg(2+) causes ΔasnB mutant cells to deform and ultimately lyse. In cell regions with deformations, PG insertion is orderly and indistinguishable from the wild-type. However, PG degradation is unevenly distributed along the sidewalls. Furthermore, ΔasnB mutant cells exhibit increased sensitivity to antibiotics targeting the cell wall. These results suggest that absence of amidated mDAP causes a lethal deregulation of PG hydrolysis that can be inhibited by increased levels of Mg(2+) . Consistently, we find that Mg(2+) inhibits autolysis of wild-type cells. We suggest that Mg(2+) helps to maintain the balance between PG synthesis and hydrolysis in cell wall mutants where this balance is perturbed in favor of increased degradation.",
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Dajkovic, A, Tesson, B, Chauhan, S, Courtin, P, Keary, R, Flores, P, Marlière, C, Filipe, SR, Chapot-Chartier, M-P & Carballido-Lopez, R 2017, 'Hydrolysis of peptidoglycan is modulated by amidation of meso-diaminopimelic acid and Mg(2+) in Bacillus subtilis', Molecular Microbiology, vol. 104, no. 6, pp. 972-988. https://doi.org/10.1111/mmi.13673

Hydrolysis of peptidoglycan is modulated by amidation of meso-diaminopimelic acid and Mg(2+) in Bacillus subtilis. / Dajkovic, Alex; Tesson, Benoit; Chauhan, Smita; Courtin, Pascal; Keary, Ruth; Flores, Pierre; Marlière, Christian; Filipe, Sérgio R; Chapot-Chartier, Marie-Pierre; Carballido-Lopez, Rut.

In: Molecular Microbiology, Vol. 104, No. 6, 24.04.2017, p. 972-988.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Hydrolysis of peptidoglycan is modulated by amidation of meso-diaminopimelic acid and Mg(2+) in Bacillus subtilis

AU - Dajkovic, Alex

AU - Tesson, Benoit

AU - Chauhan, Smita

AU - Courtin, Pascal

AU - Keary, Ruth

AU - Flores, Pierre

AU - Marlière, Christian

AU - Filipe, Sérgio R

AU - Chapot-Chartier, Marie-Pierre

AU - Carballido-Lopez, Rut

N1 - European Research Council - ERC-StG 311231 ; Marie Sklodowska Curie Individual Fellowship BACTOSHAPE - 660935

PY - 2017/4/24

Y1 - 2017/4/24

N2 - The ability of excess Mg(2+) to compensate the absence of cell wall related genes in Bacillus subtilis has been known for a long time, but the mechanism has remained obscure. Here, we show that the rigidity of wild-type cells remains unaffected with excess Mg(2+) , but the proportion of amidated meso-diaminopimelic (mDAP) acid in their peptidoglycan (PG) is significantly reduced. We identify the amidotransferase AsnB as responsible for mDAP amidation and show that the gene encoding it is essential without added Mg(2+) . Growth without excess Mg(2+) causes ΔasnB mutant cells to deform and ultimately lyse. In cell regions with deformations, PG insertion is orderly and indistinguishable from the wild-type. However, PG degradation is unevenly distributed along the sidewalls. Furthermore, ΔasnB mutant cells exhibit increased sensitivity to antibiotics targeting the cell wall. These results suggest that absence of amidated mDAP causes a lethal deregulation of PG hydrolysis that can be inhibited by increased levels of Mg(2+) . Consistently, we find that Mg(2+) inhibits autolysis of wild-type cells. We suggest that Mg(2+) helps to maintain the balance between PG synthesis and hydrolysis in cell wall mutants where this balance is perturbed in favor of increased degradation.

AB - The ability of excess Mg(2+) to compensate the absence of cell wall related genes in Bacillus subtilis has been known for a long time, but the mechanism has remained obscure. Here, we show that the rigidity of wild-type cells remains unaffected with excess Mg(2+) , but the proportion of amidated meso-diaminopimelic (mDAP) acid in their peptidoglycan (PG) is significantly reduced. We identify the amidotransferase AsnB as responsible for mDAP amidation and show that the gene encoding it is essential without added Mg(2+) . Growth without excess Mg(2+) causes ΔasnB mutant cells to deform and ultimately lyse. In cell regions with deformations, PG insertion is orderly and indistinguishable from the wild-type. However, PG degradation is unevenly distributed along the sidewalls. Furthermore, ΔasnB mutant cells exhibit increased sensitivity to antibiotics targeting the cell wall. These results suggest that absence of amidated mDAP causes a lethal deregulation of PG hydrolysis that can be inhibited by increased levels of Mg(2+) . Consistently, we find that Mg(2+) inhibits autolysis of wild-type cells. We suggest that Mg(2+) helps to maintain the balance between PG synthesis and hydrolysis in cell wall mutants where this balance is perturbed in favor of increased degradation.

KW - GRAM-POSITIVE BACTERIA

KW - WALL TEICHOIC-ACIDS

KW - SOLID-STATE NMR

KW - D-AMINO ACIDS

KW - CELL-WALL

KW - ESCHERICHIA-COLI

KW - AUTOLYTIC SYSTEM

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DO - 10.1111/mmi.13673

M3 - Article

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JO - Molecular Microbiology

JF - Molecular Microbiology

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