Identification of Novel Small Molecule Inhibitors of Twin Arginine Translocation (Tat) Pathway and Their Effect on the Control of in Chickens

Overview

Journal Front Microbiol

Specialty Microbiology

Date 2024 May 2

PMID 38694802

Authors

Loic Deblais

Mary Drozd

Anand Kumar

Janet Antwi

James Fuchs

Rahul Khupse

Yosra A Helmy

Gireesh Rajashekara

Affiliations

Soon will be listed here.

Abstract

Introduction: Control of from farm to fork is challenging due to the frequent emergence of antimicrobial-resistant isolates. Furthermore, poultry production systems are known reservoirs of . The twin-arginine translocation (Tat) pathway is a crucial bacterial secretion system that allows to colonize the host intestinal tract by using formate as the main source of energy. However, Tat pathway is also a major contributing factor for resistance to copper sulfate (CuSO).

Methods: Since mammals and chickens do not have proteins or receptors that are homologous to bacterial Tat proteins, identification of small molecule (SM) inhibitors targeting the Tat system would allow the development of safe and effective control methods to mitigate in infected or colonized hosts in both pre-harvest and post-harvest. In this study, we screened 11 commercial libraries ( = 50,917 SM) for increased susceptibility to CuSO (1 mM) in 81-176, a human isolate which is widely studied.

Results: Furthermore, we evaluated 177 SM hits (2.5 μg/mL and above) that increased the susceptibility to CuSO for the inhibition of formate dehydrogenase (Fdh) activity, a Tat-dependent substrate. Eight Tat-dependent inhibitors (T1-T8) were selected for further studies. These selected eight Tat inhibitors cleared all tested strains ( = 12) at >10 ng/mL in the presence of 0.5 mM CuSO. These selected SMs were non-toxic to colon epithelial (Caco-2) cells when treated with 50 μg/mL for 24 h and completely cleared intracellular cells when treated with 0.63 μg/mL of SM for 24 h in the presence of 0.5 mM of CuSO. Furthermore, 3 and 5-week-old chicks treated with SM candidates for 5 days had significantly decreased cecal colonization (up to 1.2 log; < 0.01) with minimal disruption of microbiota. analyses predicted that T7 has better drug-like properties than T2 inhibitor and might target a key amino acid residue (glutamine 165), which is located in the hydrophobic core of TatC protein.

Discussion: Thus, we have identified novel SM inhibitors of the Tat pathway, which represent a potential strategy to control spread on farms.

References

Kelley L, Mezulis S, Yates C, Wass M, Sternberg M . The Phyre2 web portal for protein modeling, prediction and analysis. Nat Protoc. 2015; 10(6):845-58. PMC: 5298202. DOI: 10.1038/nprot.2015.053. View

Rollauer S, Tarry M, Graham J, Jaaskelainen M, Jager F, Johnson S . Structure of the TatC core of the twin-arginine protein transport system. Nature. 2012; 492(7428):210-4. PMC: 3573685. DOI: 10.1038/nature11683. View

Connerton P, Richards P, Lafontaine G, OKane P, Ghaffar N, Cummings N . The effect of the timing of exposure to Campylobacter jejuni on the gut microbiome and inflammatory responses of broiler chickens. Microbiome. 2018; 6(1):88. PMC: 5948730. DOI: 10.1186/s40168-018-0477-5. View

Mandal S, Van Treuren W, White R, Eggesbo M, Knight R, Peddada S . Analysis of composition of microbiomes: a novel method for studying microbial composition. Microb Ecol Health Dis. 2015; 26:27663. PMC: 4450248. DOI: 10.3402/mehd.v26.27663. View

Lee M, Newell D . Campylobacter in poultry: filling an ecological niche. Avian Dis. 2006; 50(1):1-9. DOI: 10.1637/7474-111605R.1. View

Igwaran A, Okoh A . Human campylobacteriosis: A public health concern of global importance. Heliyon. 2019; 5(11):e02814. PMC: 6861584. DOI: 10.1016/j.heliyon.2019.e02814. View

Zhang , Chung , OLDENBURG . A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays. J Biomol Screen. 2000; 4(2):67-73. DOI: 10.1177/108705719900400206. View

Lee P, Tullman-Ercek D, Georgiou G . The bacterial twin-arginine translocation pathway. Annu Rev Microbiol. 2006; 60:373-95. PMC: 2654714. DOI: 10.1146/annurev.micro.60.080805.142212. View

Bageshwar U, VerPlank L, Baker D, Dong W, Hamsanathan S, Whitaker N . High Throughput Screen for Escherichia coli Twin Arginine Translocation (Tat) Inhibitors. PLoS One. 2016; 11(2):e0149659. PMC: 4764201. DOI: 10.1371/journal.pone.0149659. View

10.

Kassem I, Candelero-Rueda R, Esseili K, Rajashekara G . Formate simultaneously reduces oxidase activity and enhances respiration in Campylobacter jejuni. Sci Rep. 2017; 7:40117. PMC: 5238407. DOI: 10.1038/srep40117. View

11.

Helmy Y, Deblais L, Kassem I, Kathayat D, Rajashekara G . Novel small molecule modulators of quorum sensing in avian pathogenic Escherichia coli (APEC). Virulence. 2018; 9(1):1640-1657. PMC: 7000209. DOI: 10.1080/21505594.2018.1528844. View

12.

Tolopko A, Sullivan J, Erickson S, Wrobel D, Chiang S, Rudnicki K . Screensaver: an open source lab information management system (LIMS) for high throughput screening facilities. BMC Bioinformatics. 2010; 11:260. PMC: 3001403. DOI: 10.1186/1471-2105-11-260. View

13.

Kassem I, Zhang Q, Rajashekara G . The twin-arginine translocation system: contributions to the pathobiology of Campylobacter jejuni. Future Microbiol. 2011; 6(11):1315-27. DOI: 10.2217/fmb.11.107. View

14.

Lai C, Lai C, Lin Y, Hung C, Chu C, Feng C . Characterization of putative cholesterol recognition/interaction amino acid consensus-like motif of Campylobacter jejuni cytolethal distending toxin C. PLoS One. 2013; 8(6):e66202. PMC: 3675143. DOI: 10.1371/journal.pone.0066202. View

15.

Hall S, Hitchcock A, Butler C, Kelly D . A Multicopper oxidase (Cj1516) and a CopA homologue (Cj1161) are major components of the copper homeostasis system of Campylobacter jejuni. J Bacteriol. 2008; 190(24):8075-85. PMC: 2593206. DOI: 10.1128/JB.00821-08. View

16.

Frain K, Robinson C, van Dijl J . Transport of Folded Proteins by the Tat System. Protein J. 2019; 38(4):377-388. PMC: 6708511. DOI: 10.1007/s10930-019-09859-y. View

17.

Allos B . Campylobacter jejuni Infections: update on emerging issues and trends. Clin Infect Dis. 2001; 32(8):1201-6. DOI: 10.1086/319760. View

18.

Logue C, Sherwood J, Elijah L, Olah P, Dockter M . The incidence of Campylobacter spp. on processed turkey from processing plants in the midwestern United States. J Appl Microbiol. 2003; 95(2):234-41. DOI: 10.1046/j.1365-2672.2003.01969.x. View

19.

Lipinski C . Lead- and drug-like compounds: the rule-of-five revolution. Drug Discov Today Technol. 2014; 1(4):337-41. DOI: 10.1016/j.ddtec.2004.11.007. View

20.

Blummel A, Drepper F, Knapp B, Eimer E, Warscheid B, Muller M . Structural features of the TatC membrane protein that determine docking and insertion of a twin-arginine signal peptide. J Biol Chem. 2017; 292(52):21320-21329. PMC: 5766949. DOI: 10.1074/jbc.M117.812560. View