Dataset Information

NagA mediates the recycling of peptidoglycan derived amino sugars in mycobacteria

ABSTRACT: Many bacterial species are known to recover peptidoglycan (PG) fragments released from remodelling of their cell walls during growth and cell division. These PG fragments not only provide an essential energy resource, especially in nutrient restricted environments, but also play a critical role in influencing infection. Yet whether mycobacteria have the capacity to recycle their PG, or not, has yet to be resolved. In this study we show that NagA, an N-acetylglucosamine-6-phosphate (GlcNAc-6-P) deacetylase, is essential for coordinating the recycling of an amino sugar component released from the mycobacterial cell wall. We show that NagA is exclusively responsible for GlcNAc-6-P deacetylation and is pivotal for the de novo synthesis of core cell wall building blocks. Indeed, a nagA mutant exhibited impaired cell wall, defective biofilm formation, and enhanced susceptibility to PG targeting agents. Moreover, uptake analysis and profiling of the amino-sugar pool revealed that NagA inactivation blocks N-acetylglucosamine (GlcNAc) import and has a pronounced effect on the fate and levels of the intracellular amino sugar pool. Loss of NagA led to the up- and down-regulation of proteins involved in cell wall biosynthesis, thereby altering cell wall homeostasis. Overall, our data highlights the importance of an overlooked yet conserved component in an important PG salvage pathway in mycobacteria, in which NagA provides a unique GlcNAc sensing mechanism, thus acting as a checkpoint for regulating the recovery and reuse of PG fragments.

INSTRUMENT(S):

ORGANISM(S): Mycobacterium Smegmatis

SUBMITTER: Andrew Bottrill

LAB HEAD: Professor Elizabeth Fullam

PROVIDER: PXD065120 | Pride | 2025-11-24

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	Experiment_MS19-167_Magda_MSNAGAC1_from_OT_190710_21_MS19-167_MSNAGAC1.mzid	Mzid
	Experiment_MS19-167_Magda_MSNAGAC1_from_OT_190710_21_MS19-167_MSNAGAC1.mzid_Experiment_MS19-167_Magda_MSNAGAC1_from_OT_190710_21_MS19-167_MSNAGAC1.MGF	Mzid
	Experiment_MS19-167_Magda_MSNAGAC2_from_OT_190710_21_MS19-167_MSNAGAC2.mzid	Mzid
	Experiment_MS19-167_Magda_MSNAGAC2_from_OT_190710_21_MS19-167_MSNAGAC2.mzid_Experiment_MS19-167_Magda_MSNAGAC2_from_OT_190710_21_MS19-167_MSNAGAC2.MGF	Mzid
	Experiment_MS19-167_Magda_MSNAGAC3_from_OT_190710_21_MS19-167_MSNAGAC3.mzid	Mzid

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Publications

The deacetylase NagA mediates the remodeling and recycling of peptidoglycan-derived amino sugars in mycobacteria.

Guy Collette S CS Cooper Charlotte C Karlikowska Magdalena M Harrison James J Singh Albel A Servín-González Luis Steven LS Evans Caroline A CA Bakker Saskia E SE Bottrill Andrew A Bhatt Apoorva A Mesnage Stéphane S Besra Gurdyal S GS Fullam Elizabeth E

The Journal of biological chemistry 20250814 11

Many bacterial species are known to recover peptidoglycan (PG) fragments released from remodeling of their cell walls during growth and cell division. These PG fragments not only provide an essential energy resource, especially in nutrient restricted environments, but also play a critical role in influencing infection. Yet whether mycobacteria have the capacity to recycle their PG, or not, has still not been resolved. In this study, we show that NagA, an N-acetylglucosamine-6-phosphate (GlcNAc-6 ...[more]

PMID: 40818611

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Project description:Streptococcus pneumoniae is a human commensal bacterium that causes serious diseases. Peptidoglycan (PG) is the critical component of bacteria that maintains cell shape, size, chaining, and turgor resistance. Because of its surface exposure and eubacterial-specific mechanism of biosynthesis, PG biosynthesis remains an outstanding target for discovery of new antibiotics to resistant “superbugs,” like S. pneumoniae. Pneumococcal PG biosynthesis is carried out by a balance of septal and peripheral (side-wall-like) PG synthesis mediated by bPBP2x and bPBP2b, respectively, that emanates from midcell regions. We selected for additional suppressor mutations, besides mltG, that eliminate the requirement for essential bPBP2b in peripheral PG synthesis. We found that mutations that inactivate a putative RNA binding protein, designated KhpA, suppressed the requirement for some, but not all of the essential proteins required for peripheral PG synthesis. KhpA was used as bait in co-IP experiments to demonstrate interactions with a second RNA binding protein, designated KhpB, in cells. Mutations in khpA or khpB phenocopy each other and result in slower growth rates, smaller cell size with normal aspect ratios, and induction of cell wall stress responses. RIP-Seq analyses confirmed KhpAB as RNA binding protein in cells. Another suppressor of Δpbp2b implicated induction of a cell division operon as a way to bypass the requirement for bPBP2b. We show that increased expression of this cell division operon occurs in ΔkhpAB mutants and that this induction is necessary and sufficient to account for suppression of Δpbp2b. Comparisons of relative transcript amounts and protein levels indicate that induction of this cell division operon in ΔkhpAB mutants occurs primarily at the post-transcriptional level. Together, our results show that KhpAB, which are virulence factors in S. pneumoniae and are conserved in other Gram-positive pathogens, acts as a new class of RNA binding protein, with properties analogous to Hfq in Gram-negative bacteria.