Synergistic Effects of Anti-CmeA and Anti-CmeB Peptide Nucleic Acids on Sensitizing Campylobacter jejuni to Antibiotics

ABSTRACTThe CmeABC efflux pump inCampylobacter jejuniconfers resistance to structurally divergent antimicrobials, and inhibition of CmeABC represents a promising strategy to control antibiotic-resistantCampylobacter. Antisense peptide nucleic acids (PNAs) targeting the three components of CmeABC were evaluated for inhibition of CmeABC expression. The result revealed a synergistic effect of the PNAs targeting CmeA and CmeB on sensitizingC. jejunito antibiotics. This finding further demonstrates the feasibility of using PNAs to potentiate antibiotics against antibiotic-resistantCampylobacter.

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Inhibition of the CmeABC efflux pump by antisense peptide nucleic acids reduces the emergence of spontaneous fluoroquinolone resistant mutants in Campylobacter jejuni

10.31274/etd-20200624-17 ◽

2020 ◽

Author(s):

Kritika Singh

Keyword(s):

Nucleic Acids ◽

Campylobacter Jejuni ◽

Efflux Pump ◽

Peptide Nucleic Acids ◽

Resistant Mutants ◽

Antisense Peptide

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CmeABC Multidrug Efflux Pump Contributes to Antibiotic Resistance and Promotes Campylobacter jejuni Survival and Multiplication in Acanthamoeba polyphaga

Applied and Environmental Microbiology ◽

10.1128/aem.01600-17 ◽

2017 ◽

Vol 83 (22) ◽

Cited By ~ 11

Author(s):

Ana Vieira ◽

Amritha Ramesh ◽

Alan M. Seddon ◽

Andrey V. Karlyshev

Keyword(s):

Antibiotic Resistance ◽

Campylobacter Jejuni ◽

Molecular Mechanisms ◽

Bacterial Resistance ◽

Efflux Pump ◽

Synergistic Effects ◽

Microbial Pathogens ◽

Bacterial Survival ◽

Acanthamoeba Polyphaga ◽

Content Type

ABSTRACT Campylobacter jejuni is a foodborne pathogen that is recognized as the leading cause of human bacterial gastroenteritis. The widespread use of antibiotics in medicine and in animal husbandry has led to an increased incidence of antibiotic resistance in Campylobacter. In addition to a role in multidrug resistance (MDR), the Campylobacter CmeABC resistance-nodulation-division (RND)-type efflux pump may be involved in virulence. As a vehicle for pathogenic microorganisms, the protozoan Acanthamoeba is a good model for investigations of bacterial survival in the environment and the molecular mechanisms of pathogenicity. The interaction between C. jejuni 81-176 and Acanthamoeba polyphaga was investigated in this study by using a modified gentamicin protection assay. In addition, a possible role for the CmeABC MDR pump in this interaction was explored. Here we report that this MDR pump is beneficial for the intracellular survival and multiplication of C. jejuni in A. polyphaga but is dispensable for biofilm formation and motility. IMPORTANCE The endosymbiotic relationship between amoebae and microbial pathogens may contribute to persistence and spreading of the latter in the environment, which has significant implications for human health. In this study, we found that Campylobacter jejuni was able to survive and to multiply inside Acanthamoeba polyphaga; since these microorganisms can coexist in the same environment (e.g., on poultry farms), the latter may increase the risk of infection with Campylobacter. Our data suggest that, in addition to its role in antibiotic resistance, the CmeABC MDR efflux pump plays a role in bacterial survival within amoebae. Furthermore, we demonstrated synergistic effects of the CmeABC MDR efflux pump and TetO on bacterial resistance to tetracycline. Due to its role in both the antibiotic resistance and the virulence of C. jejuni, the CmeABC MDR efflux pump could be considered a good target for the development of antibacterial drugs against this pathogen.

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Altering behavioral responses and dopamine transporter protein with antisense peptide nucleic acids 1 1Abbreviations: DAT, dopamine transporter; and PNA, peptide nucleic acid.

Biochemical Pharmacology ◽

10.1016/s0006-2952(01)00698-0 ◽

2001 ◽

Vol 62 (7) ◽

pp. 929-932 ◽

Cited By ~ 10

Author(s):

Beth M Tyler-McMahon ◽

Jennifer A Stewart ◽

Joshua Jackson ◽

M.D Bitner ◽

Abdul Fauq ◽

...

Keyword(s):

Nucleic Acid ◽

Nucleic Acids ◽

Dopamine Transporter ◽

Peptide Nucleic Acid ◽

Peptide Nucleic Acids ◽

Behavioral Responses ◽

Transporter Protein ◽

Antisense Peptide

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Calcium Ions Effectively Enhance the Effect of Antisense Peptide Nucleic Acids Conjugated to Cationic Tat and Oligoarginine Peptides

Chemistry & Biology ◽

10.1016/j.chembiol.2005.06.009 ◽

2005 ◽

Vol 12 (8) ◽

pp. 923-929 ◽

Cited By ~ 95

Author(s):

Takehiko Shiraishi ◽

Stanislava Pankratova ◽

Peter E. Nielsen

Keyword(s):

Nucleic Acids ◽

Calcium Ions ◽

Peptide Nucleic Acids ◽

Antisense Peptide

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RNA Secondary Structure-Based Design of Antisense Peptide Nucleic Acids for Modulating Disease-Associated Aberrant Tau Pre-mRNA Alternative Splicing

Molecules ◽

10.3390/molecules24163020 ◽

2019 ◽

Vol 24 (16) ◽

pp. 3020 ◽

Cited By ~ 6

Author(s):

Alan Ann Lerk Ong ◽

Jiazi Tan ◽

Malini Bhadra ◽

Clément Dezanet ◽

Kiran M. Patil ◽

...

Keyword(s):

Alternative Splicing ◽

Nucleic Acids ◽

Splice Site ◽

Peptide Nucleic Acids ◽

Amino Sugar ◽

Hairpin Structure ◽

Mobility Shift ◽

Mobility Shift Assay ◽

Antisense Peptide ◽

Exon 10

Alternative splicing of tau pre-mRNA is regulated by a 5′ splice site (5′ss) hairpin present at the exon 10–intron 10 junction. Single mutations within the hairpin sequence alter hairpin structural stability and/or the binding of splicing factors, resulting in disease-causing aberrant splicing of exon 10. The hairpin structure contains about seven stably formed base pairs and thus may be suitable for targeting through antisense strands. Here, we used antisense peptide nucleic acids (asPNAs) to probe and target the tau pre-mRNA exon 10 5′ss hairpin structure through strand invasion. We characterized by electrophoretic mobility shift assay the binding of the designed asPNAs to model tau splice site hairpins. The relatively short (10–15 mer) asPNAs showed nanomolar binding to wild-type hairpins as well as a disease-causing mutant hairpin C+19G, albeit with reduced binding strength. Thus, the structural stabilizing effect of C+19G mutation could be revealed by asPNA binding. In addition, our cell culture minigene splicing assay data revealed that application of an asPNA targeting the 3′ arm of the hairpin resulted in an increased exon 10 inclusion level for the disease-associated mutant C+19G, probably by exposing the 5′ss as well as inhibiting the binding of protein factors to the intronic spicing silencer. On the contrary, the application of asPNAs targeting the 5′ arm of the hairpin caused an increased exon 10 exclusion for a disease-associated mutant C+14U, mainly by blocking the 5′ss. PNAs could enter cells through conjugation with amino sugar neamine or by cotransfection with minigene plasmids using a commercially available transfection reagent.

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