AsnB is responsible for peptidoglycan precursor amidation in Clostridium difficile in the presence of vancomycin

Ammam, Fariza; Patin, Delphine; Coullon, Héloïse; Blanot, Didier; Lambert, Thierry; Mengin‐Lecreulx, Dominique; Candela, Thomas

doi:10.1099/mic.0.000917

Cited by 10 publications

(15 citation statements)

References 29 publications

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“…However, while the three paralogs could restore Asn prototrophy to an E. coli asnB mutant, none was essential for producing Asn in B. subtilis, and asnB was later shown to mediate amidation of the ε-carboxyl group of mDAP residues in the peptidoglycan peptide stem (Dajkovic et al, 2017). The role of asnB homologs in mDAP amidation has also been demonstrated in other Gram-positive bacteria, including L. plantarum (Bernard et al, 2011) and C. difficile (Ammam et al, 2020). However, unlike these bacteria, which have two or three asnB paralogs, L. monocytogenes contains only one.…”

Section: Results Asnb Mutants Show Increased Sensitivity To the Antimicrobial T-cinnamaldehyde And Have Altered Cell Shapementioning

confidence: 99%

“…In the same clade, AsnB1 of L. plantarum was also demonstrated to mediate mDAP amidation (Bernard et al, 2011). The same holds for AsnB1 from C. difficile (Ammam et al, 2020), but this sequence, together with that of its AsnB2 paralog, forms a small distinct clade.…”

Section: Asnb Mediates Amidation Of Meso-diaminopimelic Acid Residues In Peptidoglycanmentioning

confidence: 86%

“…6.3.5.4). While AsnB proteins are indeed involved in Asn synthesis in several bacteria, representatives in an increasing number of Gram-positive bacteria, including L. plantarum (Bernard et al, 2011), C. glutamicum ( Levefaudes et al, 2015), B. subtilis (Dajkovic et al, 2017), and C. difficile (Ammam et al, 2020), have been shown to catalyze amidotransfer from Gln to the ε-carboxyl group of mDAP in the stem peptide of bacterial peptidoglycan rather than to Asp. Although the role of AsnB had not been studied in L. monocytogenes, there were at least two reasons to anticipate that it would also have mDAP modifying activity.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, amidation also controlled the activity of the L,D-carboxypeptidase DacB that trims the stem peptide. A similar situation -two homologs of which one specifically mediates mDAP amidation -exists in Clostridioides difficile, but a remarkable feature in this organism is that the expression of the asnB gene that confers mDAP amidation is specifically induced by vancomycin (Ammam et al, 2020). Somewhat unexpectedly, amidation slightly reduced vancomycin resistance but did not affect lysozyme resistance, and its role in C. difficile therefore remains unclear.…”

Section: Introductionmentioning

confidence: 99%

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AsnB Mediates Amidation of Meso-Diaminopimelic Acid Residues in the Peptidoglycan of Listeria monocytogenes and Affects Bacterial Surface Properties and Host Cell Invasion

et al. 2021

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A mutant of Listeria monocytogenes ScottA with a transposon in the 5' untranslated region of the asnB gene was identified to be hypersensitive to the antimicrobial t-cinnamaldehyde. Here, we report the functional characterization of AsnB in peptidoglycan (PG) modification and intracellular infection. While AsnB of Listeria is annotated as a glutamine-dependent asparagine synthase, sequence alignment showed that this protein is closely related to a subset of homologs that catalyze the amidation of meso-diaminopimelic acid (mDAP) residues in the peptidoglycan of other bacterial species. Structural analysis of peptidoglycan from an asnB mutant, compared to that of isogenic wild-type (WT) and complemented mutant strains, confirmed that AsnB mediates mDAP amidation in L. monocytogenes. Deficiency in mDAP amidation caused several peptidoglycan- and cell surface-related phenotypes in the asnB mutant, including formation of shorter but thicker cells, susceptibility to lysozyme, loss of flagellation and motility, and a strong reduction in biofilm formation. In addition, the mutant showed reduced invasion of human epithelial JEG-3 and Caco-2 cells. Analysis by immunofluorescence microscopy revealed that asnB inactivation abrogated the proper display at the listerial surface of the invasion protein InlA, which normally gets cross-linked to mDAP via its LPXTG motif. Together, this work shows that AsnB of L. monocytogenes, like several of its homologs in related Gram-positive bacteria, mediates the amidation of mDAP residues in the peptidoglycan and, in this way, affects several cell wall and cell surface-related properties. It also for the first time implicates the amidation of peptidoglycan mDAP residues in cell wall anchoring of InlA and in bacterial virulence.

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Section: Results Asnb Mutants Show Increased Sensitivity To the Antimicrobial T-cinnamaldehyde And Have Altered Cell Shapementioning

confidence: 99%

Section: Asnb Mediates Amidation Of Meso-diaminopimelic Acid Residues In Peptidoglycanmentioning

confidence: 86%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

AsnB Mediates Amidation of Meso-Diaminopimelic Acid Residues in the Peptidoglycan of Listeria monocytogenes and Affects Bacterial Surface Properties and Host Cell Invasion

et al. 2021

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“…One mechanism to confer resistance to the antibiotic vancomycin, first discovered in other Gram-positive bacteria, is to alter part of its target site through amidation of the meso -diaminopimelic acid in the peptide cross-bridge [ 12 ]. The paper in this issue looks at the function of this process in C. difficile , which the authors, led by Thomas Candela and colleagues at the Université Paris-Saclay, France, demonstrate is present in peptidoglycan isolated from C. difficile and which requires the asnB gene product [ 13 ]. Expression of this gene also appears to be induced by the presence of sub-inhibitory concentrations of vancomycin, through an unknown mechanism.…”

mentioning

confidence: 99%

Microbial Musings – June 2020

Thomas

2020

Microbiology

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Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2019–2020

Harvey

2023

Mass Spectrometry Reviews

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This review is the tenth update of the original article published in 1999 on the application of matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry to the analysis of carbohydrates and glycoconjugates and brings coverage of the literature to the end of 2020. Also included are papers that describe methods appropriate to analysis by MALDI, such as sample preparation techniques, even though the ionization method is not MALDI. The review is basically divided into three sections: (1) general aspects such as theory of the MALDI process, matrices, derivatization, MALDI imaging, fragmentation, quantification and the use of arrays. (2) Applications to various structural types such as oligo‐ and polysaccharides, glycoproteins, glycolipids, glycosides and biopharmaceuticals, and (3) other areas such as medicine, industrial processes and glycan synthesis where MALDI is extensively used. Much of the material relating to applications is presented in tabular form. The reported work shows increasing use of incorporation of new techniques such as ion mobility and the enormous impact that MALDI imaging is having. MALDI, although invented nearly 40 years ago is still an ideal technique for carbohydrate analysis and advancements in the technique and range of applications show little sign of diminishing.

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AsnB is responsible for peptidoglycan precursor amidation in Clostridium difficile in the presence of vancomycin

Abstract: Clostridium difficile 630 possesses a cryptic but functional gene cluster vanG Cd hom… Show more

Cited by 10 publications

References 29 publications

AsnB Mediates Amidation of Meso-Diaminopimelic Acid Residues in the Peptidoglycan of Listeria monocytogenes and Affects Bacterial Surface Properties and Host Cell Invasion

AsnB Mediates Amidation of Meso-Diaminopimelic Acid Residues in the Peptidoglycan of Listeria monocytogenes and Affects Bacterial Surface Properties and Host Cell Invasion

Microbial Musings – June 2020

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2019–2020

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