Structural and functional studies of pyruvate carboxylase regulation by cyclic di-AMP in lactic acid bacteria

Choi, Philip Young-Ill; Vu, Thu Minh Ngoc; Pham, Thi Huong; Woodward, Joshua J.; Turner, Mark S.; Tong, Liang

doi:10.1073/pnas.1704756114

Cited by 47 publications

(51 citation statements)

References 60 publications

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“…Considering that RePC and AnPC are evolutionarily and functionally divergent, and that the kinetic and thermodynamic features of the vast majority of PC enzymes are highly conserved, we believe that these findings are broadly applicable to all PC enzymes. Notably, a recently reported structure of PC from Lactococcus lactis revealed the BCCP domain uniquely posed in an interaction with the BC domain on a neighboring subunit (equivalent to the position occupied through translocation pathways b and c) 18 . The present study confirms and validates the catalytic relevance of this structural snapshot and extends the implications to a broad cross-section of PC enzymes.…”

Section: Discussionmentioning

confidence: 99%

Allosteric regulation alters carrier domain translocation in pyruvate carboxylase

et al. 2018

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Pyruvate carboxylase (PC) catalyzes the ATP-dependent carboxylation of pyruvate to oxaloacetate. The reaction occurs in two separate catalytic domains, coupled by the long-range translocation of a biotinylated carrier domain (BCCP). Here, we use a series of hybrid PC enzymes to examine multiple BCCP translocation pathways in PC. These studies reveal that the BCCP domain of PC adopts a wide range of translocation pathways during catalysis. Furthermore, the allosteric activator, acetyl CoA, promotes one specific intermolecular carrier domain translocation pathway. These results provide a basis for the ordered thermodynamic state and the enhanced carboxyl group transfer efficiency in the presence of acetyl CoA, and reveal that the allosteric effector regulates enzyme activity by altering carrier domain movement. Given the similarities with enzymes involved in the modular synthesis of natural products, the allosteric regulation of carrier domain movements in PC is likely to be broadly applicable to multiple important enzyme systems.

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Section: Discussionmentioning

confidence: 99%

Allosteric regulation alters carrier domain translocation in pyruvate carboxylase

et al. 2018

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“…Moreover, c-di-AMP stimulates the DNAbinding activity of the Mycobacterium smegmatis transcription factor DarR, which controls the expression of three genes (Zhang et al, 2013). Furthermore, c-di-AMP binds to and inhibits the pyruvate carboxylase in L. lactis and L. monocytogenes (Sureka et al, 2014;Choi et al, 2017). In the latter organism, c-di-AMP also binds to the cystathione-beta-synthase domain-containing (CBS) proteins CbpA and CbpB as well as to the PII-like signal transduction protein PstA (also designated as DarA) (Sureka et al, 2014;Choi et al, 2015).…”

Section: Figurementioning

confidence: 99%

An extracytoplasmic protein and a moonlighting enzyme modulate synthesis of c‐di‐AMP in Listeria monocytogenes

Gibhardt

Heidemann

Bremenkamp

et al. 2020

Environmental Microbiology

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Summary The second messenger cyclic di‐AMP (c‐di‐AMP) is essential for growth of many bacteria because it controls osmolyte homeostasis. c‐di‐AMP can regulate the synthesis of potassium uptake systems in some bacteria and it also directly inhibits and activates potassium import and export systems, respectively. Therefore, c‐di‐AMP production and degradation have to be tightly regulated depending on the environmental osmolarity. The Gram‐positive pathogen Listeria monocytogenes relies on the membrane‐bound diadenylate cyclase CdaA for c‐di‐AMP production and degrades the nucleotide with two phosphodiesterases. While the enzymes producing and degrading the dinucleotide have been reasonably well examined, the regulation of c‐di‐AMP production is not well understood yet. Here we demonstrate that the extracytoplasmic regulator CdaR interacts with CdaA via its transmembrane helix to modulate c‐di‐AMP production. Moreover, we show that the phosphoglucosamine mutase GlmM forms a complex with CdaA and inhibits the diadenylate cyclase activity in vitro. We also found that GlmM inhibits c‐di‐AMP production in L. monocytogenes when the bacteria encounter osmotic stress. Thus, GlmM is the major factor controlling the activity of CdaA in vivo. GlmM can be assigned to the class of moonlighting proteins because it is active in metabolism and adjusts the cellular turgor depending on environmental osmolarity.

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“…Accordingly, c-di-AMP synthesis was shown to be dispensable for growth on minimal media by limiting the downstream effect of the (p)ppGpp alarmone on the global regulator CodY [ 14 ]. Additionally, spontaneous mutations in pyruvate carboxylase (PycA), an enzyme of the tricarboxylic acid (TCA) cycle, also lead to a toxic accumulation of metabolites in the absence of c-di-AMP in several lactic acid bacteria [ 15 – 17 ]. However, the compensatory mechanism appears distinct in other bacteria.…”

Section: Introductionmentioning

confidence: 99%

Cyclic di-AMP regulation of osmotic homeostasis is essential in Group B Streptococcus

et al. 2018

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Cyclic nucleotides are universally used as secondary messengers to control cellular physiology. Among these signalling molecules, cyclic di-adenosine monophosphate (c-di-AMP) is a specific bacterial second messenger recognized by host cells during infections and its synthesis is assumed to be necessary for bacterial growth by controlling a conserved and essential cellular function. In this study, we sought to identify the main c-di-AMP dependent pathway in Streptococcus agalactiae, the etiological agent of neonatal septicaemia and meningitis. By conditionally inactivating dacA, the only diadenyate cyclase gene, we confirm that c-di-AMP synthesis is essential in standard growth conditions. However, c-di-AMP synthesis becomes rapidly dispensable due to the accumulation of compensatory mutations. We identified several mutations restoring the viability of a ΔdacA mutant, in particular a loss-of-function mutation in the osmoprotectant transporter BusAB. Identification of c-di-AMP binding proteins revealed a conserved set of potassium and osmolyte transporters, as well as the BusR transcriptional factor. We showed that BusR negatively regulates busAB transcription by direct binding to the busAB promoter. Loss of BusR repression leads to a toxic busAB expression in absence of c-di-AMP if osmoprotectants, such as glycine betaine, are present in the medium. In contrast, deletion of the gdpP c-di-AMP phosphodiesterase leads to hyperosmotic susceptibility, a phenotype dependent on a functional BusR. Taken together, we demonstrate that c-di-AMP is essential for osmotic homeostasis and that the predominant mechanism is dependent on the c-di-AMP binding transcriptional factor BusR. The regulation of osmotic homeostasis is likely the conserved and essential function of c-di-AMP, but each species has evolved specific c-di-AMP mechanisms of osmoregulation to adapt to its environment.

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Structural and functional studies of pyruvate carboxylase regulation by cyclic di-AMP in lactic acid bacteria

Cited by 47 publications

References 60 publications

Allosteric regulation alters carrier domain translocation in pyruvate carboxylase

Allosteric regulation alters carrier domain translocation in pyruvate carboxylase

An extracytoplasmic protein and a moonlighting enzyme modulate synthesis of c‐di‐AMP in Listeria monocytogenes

Cyclic di-AMP regulation of osmotic homeostasis is essential in Group B Streptococcus

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