Type II Fatty Acid Synthesis Is Essential for the Replication of Chlamydia trachomatis

Yao, Jiangwei; AbdelRahman, Yasser M.; Robertson, R.M.; Cox, John V.; Belland, Robert J.; White, Stephen W.; Rock, Charles O.

doi:10.1074/jbc.m114.584185

Cited by 44 publications

(69 citation statements)

References 48 publications

(63 reference statements)

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“…A NdeI restriction site was engineered at the 5Ј-end of the gene with the start codon in the NdeI site, whereas a His 6 tag, stop codon, and an EcoRI restriction site were sequentially engineered at the 3Ј-end of the gene. The genes were cloned into the pPJ131 plasmid (a modified version of the pBluescript plasmid with the multiple cloning site from pET28a) and the pET21a plasmid via the NdeI and EcoRI (New England Biolabs) restriction sites (24).…”

Section: Methodsmentioning

confidence: 99%

“…Cells were pelleted and washed twice with 20 mM Tris, pH 8.2, and finally resuspended in 20 mM Tris, pH 8.2 (20 ml/liter of culture). Cells were lysed via a cell disruptor and the C-terminal His 6 -tagged NgFabI was purified via standard nickel chelation chromatography (24).…”

Section: Methodsmentioning

confidence: 99%

“…The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. 1 enoyl-ACP reductase (FabI) (23)(24)(25)(26). FabI catalyzes the reduction of trans-2-enoyl-ACP, a rate-controlling step required to complete each round of elongation in fatty acid synthesis (27 …”

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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: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

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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“…Therefore, viewing AasC as an opportunistic pathway with a role in increasing the overall cellular yield of C. trachomatis by sparing energy for other biosynthetic pathways is consistent with the bacterial paradigm. Endogenous C. trachomatis fatty acid synthesis is essential for the replication of C. trachomatis on the basis of the blockade of growth by a FASII inhibitor (9). However, this fact alone cannot be used to conclude that C. trachomatis does not support its growth with host fatty acids.…”

Section: Discussionmentioning

confidence: 99%

“…Autonomous phospholipid synthesis is shown. The fatty acid synthesis module (FASII) supplies acyl-ACP to the PlsE and PlsC acyltransferases of phospholipid biosynthesis and to the lipid A acyltransferases that initiate lipo-oligosaccharide (LOS) biosynthesis, as described previously (6,9). The new enzymes characterized in this work are shown in light blue.…”

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confidence: 95%

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

Yao

Dodson

Frank

et al. 2015

Journal of Biological Chemistry

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Background: C. trachomatis relies on its own biosynthetic pathways to produce membrane phospholipids. Results: C. trachomatis expresses an acyl-acyl carrier protein synthetase to activate fatty acids. Conclusion: C. trachomatis utilizes fatty acids obtained from the host to construct phospholipids. Significance: C. trachomatis selectively scavenges host saturated fatty acids, the most energy-expensive component needed for phospholipid synthesis.

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The Molecular Basis of Catalysis by SDR Family Members Ketoacyl‐ACP Reductase FabG and Enoyl‐ACP Reductase FabI in Type‐II Fatty Acid Biosynthesis

Zhou,

Zhang,

Wang

et al. 2023

Angewandte Chemie

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The short‐chain dehydrogenase/reductase (SDR) superfamily members acyl‐ACP reductases FabG and FabI are indispensable core enzymatic modules and catalytic orientation controllers in type‐II fatty acid biosynthesis. Herein, we report their distinct substrate allosteric recognition and enantioselective reduction mechanisms. FabG achieves allosteric regulation of ACP and NADPH through ACP binding across two adjacent FabG monomers, while FabI follows an irreversible compulsory order of substrate binding in that NADH binding must precede that of ACP on a discrete FabI monomer. Moreover, FabG and FabI utilize a backdoor residue Phe187 or a “rheostat” α8 helix for acyl chain length selection, and their corresponding triad residues Ser142 or Tyr145 recognize the keto‐ or enoyl‐acyl substrates, respectively, facilitating initiation of nucleophilic attack by NAD(P)H. The other two triad residues (Tyr and Lys) mediate subsequent proton transfer and (R)‐3‐hydroxyacyl‐ or saturated acyl‐ACP production.

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Type II Fatty Acid Synthesis Is Essential for the Replication of Chlamydia trachomatis

Cited by 44 publications

References 48 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

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

The Molecular Basis of Catalysis by SDR Family Members Ketoacyl‐ACP Reductase FabG and Enoyl‐ACP Reductase FabI in Type‐II Fatty Acid Biosynthesis

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