Citation

BibTex format

@article{Tosi:2019:10.1371/journal.ppat.1007537,
author = {Tosi, T and Hoshiga, F and Millership, C and Singh, R and Eldrid, C and Patin, D and Mengin-Lecreulx, D and Thalassinos, K and Freemont, P and Grundling, A},
doi = {10.1371/journal.ppat.1007537},
journal = {PLoS Pathogens},
pages = {1--28},
title = {Inhibition of the Staphylococcus aureus c-di-AMP cyclase DacA by direct interaction with the phosphoglucosamine mutase GlmM},
url = {http://dx.doi.org/10.1371/journal.ppat.1007537},
volume = {15},
year = {2019}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - c-di-AMP is an important second messenger molecule that plays a pivotal role in regulating fundamental cellular processes, including osmotic and cell wall homeostasis in many Gram-positive organisms. In the opportunistic human pathogen Staphylococcus aureus, c-di-AMP is produced by the membrane-anchored DacA enzyme. Inactivation of this enzyme leads to a growth arrest under standard laboratory growth conditions and a re-sensitization of methicillin-resistant S. aureus (MRSA) strains to ß-lactam antibiotics. The gene coding for DacA is part of the conserved three-gene dacA/ybbR/glmM operon that also encodes the proposed DacA regulator YbbR and the essential phosphoglucosamine mutase GlmM, which is required for the production of glucosamine-1-phosphate, an early intermediate of peptidoglycan synthesis. These three proteins are thought to form a complex in vivo and, in this manner, help to fine-tune the cellular c-di-AMP levels. To further characterize this important regulatory complex, we conducted a comprehensive structural and functional analysis of the S. aureus DacA and GlmM enzymes by determining the structures of the S. aureus GlmM enzyme and the catalytic domain of DacA. Both proteins were found to be dimers in solution as well as in the crystal structures. Further site-directed mutagenesis, structural and enzymatic studies showed that multiple DacA dimers need to interact for enzymatic activity. We also show that DacA and GlmM form a stable complex in vitro and that S. aureus GlmM, but not Escherichia coli or Pseudomonas aeruginosa GlmM, acts as a strong inhibitor of DacA function without the requirement of any additional cellular factor. Based on Small Angle X-ray Scattering (SAXS) data, a model of the complex revealed that GlmM likely inhibits DacA by masking the active site of the cyclase and preventing higher oligomer formation. Together these results provide an important mechanistic insight into how c-di-AMP production can be regulated in the cell.
AU  - Tosi,T
AU  - Hoshiga,F
AU  - Millership,C
AU  - Singh,R
AU  - Eldrid,C
AU  - Patin,D
AU  - Mengin-Lecreulx,D
AU  - Thalassinos,K
AU  - Freemont,P
AU  - Grundling,A
DO  - 10.1371/journal.ppat.1007537
EP  - 28
PY  - 2019///
SN  - 1553-7366
SP  - 1
TI  - Inhibition of the Staphylococcus aureus c-di-AMP cyclase DacA by direct interaction with the phosphoglucosamine mutase GlmM
T2  - PLoS Pathogens
UR  - http://dx.doi.org/10.1371/journal.ppat.1007537
UR  - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000457400000035&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR  - https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1007537
UR  - http://hdl.handle.net/10044/1/66840
VL  - 15
ER  -
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