Chlamydia trachomatis Scavenges Host Fatty Acids for Phospholipid Synthesis via an Acyl-Acyl Carrier Protein Synthetase

Yao, Jiangwei; Dodson, V. Joshua; Frank, Matthew W.; Rock, Charles O.

doi:10.1074/jbc.m115.671008

Cited by 38 publications

(62 citation statements)

References 44 publications

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“…In contrast, strain LCH30 expressing the C. trachomatis acyl-ACP synthetase (aasC) converted D3-14:0 to D3-16:0 prior to incorporation into phospholipid (Fig. 5C), consistent with the formation of acyl-ACP that can either be used for phospholipid synthesis or enter the FASII elongation cycle (49). Strain LCH30 expressing NGO0530 incorporated D3-14:0 both as D3-14:0 and elongated D3-14:0 to D3-16:0, demonstrating that NGO0530 functioned as an acyl-ACP synthetase (Fig.…”

Section: Incorporation Of Exogenous Fatty Acids Into Phospholipids-mentioning

confidence: 81%

“…Vibrio harveyi expresses an acyl-ACP synthetase capable of releasing acyl-ACP that is used by FASII or the PlsB/PlsC acyltransferases (59,60). C. trachomatis also expresses an acyl-ACP synthetase that produces acyl-ACP for FASII and the PlsE/PlsC acyltransferase system (49). The function(s) for an acyl-CoA synthetase in N. gonorrhoeae is not clear.…”

Section: Discussionmentioning

confidence: 99%

“…Characterization of NGO0530 and NGO1213-N. gonorrhoeae have two genes (NGO0530 and NGO1213) that encode members of the superfamily of proteins containing the AFD_class_I domain, which corresponds to the binding site for the acyl-adenylate intermediate in acyl-ACP and acyl-CoA synthetases (12,49). The functions of the two potential acyl-CoA/ACP synthetase genes were determined by their expression in E. coli and isotopic fatty acid labeling.…”

Section: Incorporation Of Exogenous Fatty Acids Into Phospholipids-mentioning

confidence: 99%

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Activation of Exogenous Fatty Acids to Acyl-Acyl Carrier Protein Cannot Bypass FabI Inhibition in Neisseria

Yao

Bruhn

Frank

et al. 2016

Journal of Biological Chemistry

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Neisseria is a Gram-negative pathogen with phospholipids composed of straight chain saturated and monounsaturated fatty acids, the ability to incorporate exogenous fatty acids, and lipopolysaccharides that are not essential. The FabI inhibitor, AFN-1252, was deployed as a chemical biology tool to determine whether Neisseria can bypass the inhibition of fatty acid synthesis by incorporating exogenous fatty acids. Neisseria encodes a functional FabI that was potently inhibited by AFN-1252. AFN-1252 caused a dose-dependent inhibition of fatty acid synthesis in growing Neisseria, a delayed inhibition of growth phenotype, and minimal inhibition of DNA, RNA, and protein synthesis, showing that its mode of action is through inhibiting fatty acid synthesis. Isotopic fatty acid labeling experiments showed that Neisseria encodes the ability to incorporate exogenous fatty acids into its phospholipids by an acyl-acyl carrier protein-dependent pathway. However, AFN-1252 remained an effective antibacterial when Neisseria were supplemented with exogenous fatty acids. These results demonstrate that extracellular fatty acids are activated by an acyl-acyl carrier protein synthetase (AasN) and validate type II fatty acid synthesis (FabI) as a therapeutic target against Neisseria.Neisseria is a genus of mucosal colonizing bacteria found in a variety of animals. Of the 11 species of Neisseria associated with humans, Neisseria gonorrhoeae and Neisseria meningitidis are obligate human pathogens. N. meningitidis is a common cause of meningitis and other meningococcal diseases in infants and children with 3,000 cases/year in the United States (1). N. gonorrhoeae is the causative agent of the sexually transmitted disease gonorrhoeae, with Ͼ100 million new cases/year worldwide (2). The incidence of multidrug-resistant N. gonorrhoeae has markedly increased in recent years, raising the prospect of untreatable gonorrhoeae and highlighting the need for developing new antibiotics and discovering new antibiotic targets in Neisseria (2). One emerging antibiotic drug target is bacterial type II fatty acid synthesis (FASII) 3 (3-6). All characterized bacteria have the ability to utilize exogenous fatty acids for phospholipid synthesis (7); thus, it is important to understand the pathways for fatty acid uptake and whether extracellular fatty acids can bypass FASII inhibition (7-9).Neisseria are Gram-negative bacteria with phospholipids containing saturated and unsaturated fatty acids (10) and LPS containing 3-hydroxy fatty acids (11). Escherichia coli utilizes an acyl-CoA synthetase (FadD) to activate exogenous fatty acids (12), and they are incorporated into phospholipids by the PlsB/PlsC acyltransferase system that utilizes either acyl-ACP or acyl-CoA (7, 13). E. coli cannot bypass FASII inhibitors because there is no mechanism for the generation of acyl-ACP from extracellular fatty acids, and FASII is the only source for the 3-hydroxy fatty acids required to synthesize the essential LPS (14, 15). The fatty acid composition of Neisseria resembl...

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Section: Incorporation Of Exogenous Fatty Acids Into Phospholipids-mentioning

confidence: 81%

Section: Discussionmentioning

confidence: 99%

Section: Incorporation Of Exogenous Fatty Acids Into Phospholipids-mentioning

confidence: 99%

See 1 more Smart Citation

Activation of Exogenous Fatty Acids to Acyl-Acyl Carrier Protein Cannot Bypass FabI Inhibition in Neisseria

Yao

Bruhn

Frank

et al. 2016

Journal of Biological Chemistry

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“…Due to differences between FASII and the monofunctional mammalian type I fatty acid synthase [3], targeting FASII enzymes for novel antibiotic therapeutics is an active area of research [4–7]. However, all bacteria characterized to date can assimilate exogenous fatty acids [8–12], but only some species can bypass inhibition of FASII by incorporating exogenous fatty acids [13]. Several FASII inhibitors are currently in clinical development [14–20], making understanding the mechanism of exogenous fatty acid activation for phospholipid synthesis and whether exogenous fatty acids can bypass FASII inhibition in pathogenic bacterial species an important area of research.…”

Section: Introductionmentioning

confidence: 99%

Exogenous fatty acid metabolism in bacteria

2017

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Bacterial type II fatty acid synthesis (FASII) is a target for novel antibiotic development. All bacteria encode for mechanisms to incorporate exogenous fatty acids, and some bacteria can use exogenous fatty acids to bypass FASII inhibition. Bacteria encode three different mechanisms for activating exogenous fatty acids for incorporation into phospholipid synthesis. Exogenous fatty acids are converted into acyl-CoA in Gammaproteobacteria such as E. coli. Acyl-CoA molecules constitute a separate pool from endogenously synthesized acyl-ACP. Acyl-CoA can be used for phospholipid synthesis or broken down by β-oxidation, but cannot be used for lipopolysaccharide synthesis. Exogenous fatty acids are converted into acyl-ACP in some Gram-negative bacteria. The resulting acyl-ACP undergoes the same fates as endogenously synthesized acyl-ACP. Exogenous fatty acids are converted into acyl-phosphates in Gram-positive bacteria, and can be used for phospholipid synthesis or become acyl-ACP. Only the order Lactobacillales can use exogenous fatty acids to bypass FASII inhibition. FASII shuts down completely in presence of exogenous fatty acids in Lactobacillales, allowing Lactobacillales to synthesize phospholipids entirely from exogenous fatty acids. Inhibition of FASII cannot be bypassed in other bacteria because FASII is only partially down-regulated in presence of exogenous fatty acid or FASII is required to synthesize essential metabolites such as β-hydroxyacyl-ACP. Certain selective pressures such as FASII inhibition or growth in biofilms can select for naturally occurring one step mutations that attenuate endogenous fatty acid synthesis. Although attempts have been made to estimate the natural prevalence of these mutants, culture-independent metagenomic methods would provide a better estimate.

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“…The unique aspect of L. monocytogenes is that the L. monocytogenes genome has four putative enoyl-acyl carrier protein reductase genes of three different isoforms. (25), and analyzed via molecular species analysis to determine the phospholipid species (24,26). Growth curves and molecular species analysis experiments were conducted on two independent biological replicates, and representative curves and spectra are shown in the figures.…”

Section: Methodsmentioning

confidence: 99%

Enoyl-Acyl Carrier Protein Reductase I (FabI) Is Essential for the Intracellular Growth of Listeria monocytogenes

et al. 2016

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Listeria monocytogenes is a facultative intracellular Gram-positive bacterium that is the primary cause of the disease listeriosis (1). The environmentally hardy and ubiquitous L. monocytogenes is ingested through contaminated food products and can cause noninvasive diseases, such as gastroenteritis, or invade through the intestinal epithelium to cause central nervous system infections or bacteremia. L. monocytogenes poses a major medical threat for pregnant women and immunocompromised patients. Yearly outbreaks of L. monocytogenes in a variety of food products from fruits to ice cream underscore the need to understand the requirements for L. monocytogenes infection and discover new methods to inhibit the growth of L. monocytogenes.The phospholipids of L. monocytogenes are composed of branched-chain fatty acids synthesized by the L. monocytogenes type II fatty acid synthesis system. Decreasing the synthesis of branchedchain fatty acids by the genetic deletion of the branched-chain ␣-keto acid dehydrogenase compromises the environmental survival and intracellular pathogenesis of L. monocytogenes (2-6). The pathways for fatty acid synthesis, phospholipid synthesis, and exogenous fatty acid incorporation of L. monocytogenes are predicted to be the same as those for the phylogenetically related and wellcharacterized Staphylococcus aureus system (Fig. 1). Fatty acids are synthesized using a prototypical type II bacterial fatty acid synthesis, with 2-methylbutyryl-coenzyme A (CoA) as the primer, to make branch-chain fatty acids (3,5,(7)(8)(9)). An acyl-acyl carrier protein (ACP) of the appropriate length is used as the acyl donor by the PlsX/Y/C system to make phosphatidic acid from glycerol-3-phosphate (10, 11). Exogenous fatty acids are incorporated via the fatty acid kinase system (12)(13)(14). An unusual feature of L. monocytogenes fatty acid synthesis is the presence of four genes that may encode enoyl-acyl carrier protein reductases, including a FabI, a FabL, and two FabK isoforms, rather than the single FabI found in S. aureus and a variety of other bacterial pathogens. The FabL isoform was discovered in the phylogenetically related Bacillus, which also has a FabI (15). Both proteins function in fatty acid synthesis in Bacillus. The FabK isoform is a flavoprotein unrelated to FabI that was originally discovered in Streptococcus pneumoniae (16). The Enterococcus faecalis genome encodes both FabI and FabK, but FabI is responsible for the bulk of fatty acid synthesis while FabK plays a minor role in supporting fatty acid synthesis (17).The goal of this study was to identify which of the putative L. monocytogenes reductase genes code for functional enoyl-acyl carrier protein reductases, assess their contribution to supporting type II bacterial fatty acid synthesis, and determine their role in planktonic and intracellular growth. Complete inhibition of FabI using the FabI selective inhibitor AFN-1252 reduced the fatty acid synthesis of L. monocytogenes strain 10403S by 80% and reduced the growth rate of L. monocytog...

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Chlamydia trachomatis Scavenges Host Fatty Acids for Phospholipid Synthesis via an Acyl-Acyl Carrier Protein Synthetase

Cited by 38 publications

References 44 publications

Activation of Exogenous Fatty Acids to Acyl-Acyl Carrier Protein Cannot Bypass FabI Inhibition in Neisseria

Activation of Exogenous Fatty Acids to Acyl-Acyl Carrier Protein Cannot Bypass FabI Inhibition in Neisseria

Exogenous fatty acid metabolism in bacteria

Enoyl-Acyl Carrier Protein Reductase I (FabI) Is Essential for the Intracellular Growth of Listeria monocytogenes

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