Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Proteomics-Based RT-qPCR and Functional Analysis of 18 Genes in Metronidazole Resistance of Bacteroides fragilis

Bakhtiyar Mahmood
ORCID logo ,
Anna Paunkov
,
Malgorzata Kupc
,
Katalin Burián
ORCID logo ,
Erzsébet Nagy
,
David Leitsch
,
József Sóki
* ORCID logo
Version 1 : Received: 17 January 2024 / Approved: 17 January 2024 / Online: 17 January 2024 (13:06:47 CET)

A peer-reviewed article of this Preprint also exists.

Mahmood, B.; Paunkov, A.; Kupc, M.; Burián, K.; Nagy, E.; Leitsch, D.; Sóki, J. Proteomics-Based RT-qPCR and Functional Analysis of 18 Genes in Metronidazole Resistance of Bacteroides fragilis. Antibiotics 2024, 13, 207. Mahmood, B.; Paunkov, A.; Kupc, M.; Burián, K.; Nagy, E.; Leitsch, D.; Sóki, J. Proteomics-Based RT-qPCR and Functional Analysis of 18 Genes in Metronidazole Resistance of Bacteroides fragilis. Antibiotics 2024, 13, 207.

Abstract

Previously, we reported that metronidazole MICs are not dependent on the expression levels of nim genes in B. fragilis strains; and we also compared the proteomes of metronidazole-resistant laboratory B. fragilis strains to those of their susceptible parent strains. Here, we used RT-qPCR to correlate the expression levels of 18 candidate genes in a panel of selected, clinical nim gene-positive and -negative B. fragilis strains to their metronidazole MICs. Metronidazole MICs were correlated to the expression of certain tested genes. Specifically, lactate dehydrogenase expression correlated positively, whereas cytochrome fumarate reductase/succinate dehydrogenase, malate dehydrogenase, phosphoglycerate kinase redox and gat (GCN5-like acetyltransferase), and relA (stringent response) regulatory gene expressions correlated negatively with metronidazole MICs. This result provides evidence for the involvement of carbohydrate catabolic enzymes in metronidazole resistance in B. fragilis. This result was supported by direct substrate utilization tests. However, the exact roles of these genes/proteins should be determined in deletion–complementation tests. Moreover, the exact redox cofactor(s) participating in metronidazole activation need to be identified.

Keywords

Bacteroides, metronidazole, nim, resistance mechanism, RT-qPCR

Subject

Biology and Life Sciences, Immunology and Microbiology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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