Cyclic-di-AMP synthesis by the diadenylate cyclase CdaA is modulated by the peptidoglycan biosynthesis enzyme GlmM in lactococcus lactis

Zhu, Y; Pham, TH; Nhiep, THN; Vu, NMT; Marcellin, E; Chakrabortti, A; Wang, Y; Waanders, J; Lo, R; Huston, WM; Bansal, N; Nielsen, LK; Liang, ZX; Turner, MS

Cyclic-di-AMP synthesis by the diadenylate cyclase CdaA is modulated by the peptidoglycan biosynthesis enzyme GlmM in lactococcus lactis

Zhu, Y Pham, TH Nhiep, THN Vu, NMT Marcellin, E Chakrabortti, A Wang, Y Waanders, J Lo, R Huston, WM

Bansal, N Nielsen, LK Liang, ZX Turner, MS

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Publication Type:: Journal Article
Citation:: Molecular Microbiology, 2016, 99 (6), pp. 1015 - 1027
Issue Date:: 2016-03-01

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Field	Value	Language
dc.contributor.author	Zhu, Y	en_US
dc.contributor.author	Pham, TH	en_US
dc.contributor.author	Nhiep, THN	en_US
dc.contributor.author	Vu, NMT	en_US
dc.contributor.author	Marcellin, E	en_US
dc.contributor.author	Chakrabortti, A	en_US
dc.contributor.author	Wang, Y	en_US
dc.contributor.author	Waanders, J	en_US
dc.contributor.author	Lo, R	en_US
dc.contributor.author	Huston, WM https://orcid.org/0000-0002-0879-1287	en_US
dc.contributor.author	Bansal, N	en_US
dc.contributor.author	Nielsen, LK	en_US
dc.contributor.author	Liang, ZX	en_US
dc.contributor.author	Turner, MS	en_US
dc.date.available	2020-05-25T19:03:08Z
dc.date.issued	2016-03-01	en_US
dc.identifier.citation	Molecular Microbiology, 2016, 99 (6), pp. 1015 - 1027	en_US
dc.identifier.issn	0950-382X	en_US
dc.identifier.uri	http://hdl.handle.net/10453/41116
dc.description.abstract	© 2016 John Wiley & Sons Ltd. The second messenger cyclic-di-adenosine monophosphate (c-di-AMP) plays important roles in growth, virulence, cell wall homeostasis, potassium transport and affects resistance to antibiotics, heat and osmotic stress. Most Firmicutes contain only one c-di-AMP synthesizing diadenylate cyclase (CdaA); however, little is known about signals and effectors controlling CdaA activity and c-di-AMP levels. In this study, a genetic screen was employed to identify components which affect the c-di-AMP level in Lactococcus. We characterized suppressor mutations that restored osmoresistance to spontaneous c-di-AMP phosphodiesterase gdpP mutants, which contain high c-di-AMP levels. Loss-of-function and gain-of-function mutations were identified in the cdaA and gdpP genes, respectively, which led to lower c-di-AMP levels. A mutation was also identified in the phosphoglucosamine mutase gene glmM, which is commonly located within the cdaA operon in bacteria. The glmM I154F mutation resulted in a lowering of the c-di-AMP level and a reduction in the key peptidoglycan precursor UDP-N-acetylglucosamine in L. lactis. C-di-AMP synthesis by CdaA was shown to be inhibited by GlmMI154F more than GlmM and GlmMI154F was found to bind more strongly to CdaA than GlmM. These findings identify GlmM as a c-di-AMP level modulating protein and provide a direct connection between c-di-AMP synthesis and peptidoglycan biosynthesis. c-di-AMP is an essential signalling molecule which affects peptidoglycan homeostasis and resistance against various stressors, however little is known regarding how the c-di-AMP level is regulated in the cell. Here we identify the peptidoglycan biosynthesis enzyme GlmM as a modulator of c-di-AMP synthesis through its regulation of diadenylate cyclase enzyme CdaA activity in Lactococcus lactis.	en_US
dc.relation.ispartof	Molecular Microbiology	en_US
dc.relation.isbasedon	10.1111/mmi.13281	en_US
dc.subject.classification	Microbiology	en_US
dc.subject.mesh	Cell Wall	en_US
dc.subject.mesh	Lactococcus lactis	en_US
dc.subject.mesh	Phosphoric Diester Hydrolases	en_US
dc.subject.mesh	Phosphoglucomutase	en_US
dc.subject.mesh	Phosphorus-Oxygen Lyases	en_US
dc.subject.mesh	Peptidoglycan	en_US
dc.subject.mesh	Bacterial Proteins	en_US
dc.subject.mesh	Dinucleoside Phosphates	en_US
dc.subject.mesh	Cyclic AMP	en_US
dc.subject.mesh	Adenosine Monophosphate	en_US
dc.subject.mesh	Second Messenger Systems	en_US
dc.subject.mesh	Adenylyl Cyclases	en_US
dc.title	Cyclic-di-AMP synthesis by the diadenylate cyclase CdaA is modulated by the peptidoglycan biosynthesis enzyme GlmM in lactococcus lactis	en_US
dc.type	Journal Article
utslib.citation.volume	6	en_US
utslib.citation.volume	99	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	06 Biological Sciences	en_US
utslib.for	07 Agricultural and Veterinary Sciences	en_US
utslib.for	11 Medical and Health Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	open_access
pubs.issue	6	en_US
pubs.publication-status	Published	en_US
pubs.volume	99	en_US

Abstract:

© 2016 John Wiley & Sons Ltd. The second messenger cyclic-di-adenosine monophosphate (c-di-AMP) plays important roles in growth, virulence, cell wall homeostasis, potassium transport and affects resistance to antibiotics, heat and osmotic stress. Most Firmicutes contain only one c-di-AMP synthesizing diadenylate cyclase (CdaA); however, little is known about signals and effectors controlling CdaA activity and c-di-AMP levels. In this study, a genetic screen was employed to identify components which affect the c-di-AMP level in Lactococcus. We characterized suppressor mutations that restored osmoresistance to spontaneous c-di-AMP phosphodiesterase gdpP mutants, which contain high c-di-AMP levels. Loss-of-function and gain-of-function mutations were identified in the cdaA and gdpP genes, respectively, which led to lower c-di-AMP levels. A mutation was also identified in the phosphoglucosamine mutase gene glmM, which is commonly located within the cdaA operon in bacteria. The glmM I154F mutation resulted in a lowering of the c-di-AMP level and a reduction in the key peptidoglycan precursor UDP-N-acetylglucosamine in L. lactis. C-di-AMP synthesis by CdaA was shown to be inhibited by GlmMI154F more than GlmM and GlmMI154F was found to bind more strongly to CdaA than GlmM. These findings identify GlmM as a c-di-AMP level modulating protein and provide a direct connection between c-di-AMP synthesis and peptidoglycan biosynthesis. c-di-AMP is an essential signalling molecule which affects peptidoglycan homeostasis and resistance against various stressors, however little is known regarding how the c-di-AMP level is regulated in the cell. Here we identify the peptidoglycan biosynthesis enzyme GlmM as a modulator of c-di-AMP synthesis through its regulation of diadenylate cyclase enzyme CdaA activity in Lactococcus lactis.

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http://hdl.handle.net/10453/41116