A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression
Authors
Ćurčić, JovanaJakovljević, Stefan
Novović, Katarina
Vasiljević, Zorica
Kojić, Milan
Jovčić, Branko
Malešević, Milka
Conference object (Published version)
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Introduction: Quorum quenching (QQ) isthe enzymatic degradation of cell-to-cellsignaling molecules.
In this study, the potential of the novel YtnP lactonase, the quorum quenching enzyme derived from S.
maltophilia, to reduce P. aeruginosa quorum sensing and virulence factor gene expression was investigated.
Methods: MMA83 culture (adjusted to 1.5x105 CFU/ml) was treated with recombinant YtnP lactonase
(final concentration 50 μg/ml) at 37°C for 12 hours under aeration. RNA isolation of the treated and untreated MMA83 culture was performed using the RNeasy Mini Kit (Qiagen, Germany) according to the
protocol. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR), was used to analyze
the effect ofYtnP lactonase on the relative mRNA levels of the LasI/LasR, RhiI/RhiR, and PQS signaling network genes of P. aeruginosa MMA83 and virulence factor genes. The rpsL was used as an endogenous
control to normalize obtained data following the 2-ΔΔCt method.
Results: The QS ge...nes belonging to three QS networks – LasI/LasR, RhiI/RhiR, and PQS of P. aeruginosa
MMA83 treated with YtnP lactonase were significantly downregulated. The RT -qPCR results show that
treatment with YtnP-lactonase decreased the relative mRNA levels of genes involved in the production
of elastase (lasB approximately 2-fold), alginate (algK approximately 2.2-fold), pyocyanin (phzM approximately 3.5-fold), pyoverdin (pvdS approximately 2-fold), and rhamnolipid (rhlC approximately 4-fold).
These results suggest that YtnP lactonase exerts an antivirulence effect at the transcription level.
Conclusion: YtnP lactonase, a quorum quenching (QQ) enzyme, has the potential to be used as an innovative enzyme-based antivirulence therapeutic to combat infections caused by P. aeruginosa.
Keywords:
Stenotrophomonas maltophilia / lactonases, antivirulence therapy / Pseudomonas aeruginosa / quorum sensingSource:
CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia, 2023, 121-121Publisher:
- Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
Funding / projects:
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200042 (University of Belgrade, Institute of Molecular Genetics and Genetic Engineering) (RS-MESTD-inst-2020-200042)
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200178 (University of Belgrade, Faculty of Biology) (RS-MESTD-inst-2020-200178)
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TorlakTY - CONF AU - Ćurčić, Jovana AU - Jakovljević, Stefan AU - Novović, Katarina AU - Vasiljević, Zorica AU - Kojić, Milan AU - Jovčić, Branko AU - Malešević, Milka PY - 2023 UR - http://intor.torlakinstitut.com/handle/123456789/803 AB - Introduction: Quorum quenching (QQ) isthe enzymatic degradation of cell-to-cellsignaling molecules. In this study, the potential of the novel YtnP lactonase, the quorum quenching enzyme derived from S. maltophilia, to reduce P. aeruginosa quorum sensing and virulence factor gene expression was investigated. Methods: MMA83 culture (adjusted to 1.5x105 CFU/ml) was treated with recombinant YtnP lactonase (final concentration 50 μg/ml) at 37°C for 12 hours under aeration. RNA isolation of the treated and untreated MMA83 culture was performed using the RNeasy Mini Kit (Qiagen, Germany) according to the protocol. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR), was used to analyze the effect ofYtnP lactonase on the relative mRNA levels of the LasI/LasR, RhiI/RhiR, and PQS signaling network genes of P. aeruginosa MMA83 and virulence factor genes. The rpsL was used as an endogenous control to normalize obtained data following the 2-ΔΔCt method. Results: The QS genes belonging to three QS networks – LasI/LasR, RhiI/RhiR, and PQS of P. aeruginosa MMA83 treated with YtnP lactonase were significantly downregulated. The RT -qPCR results show that treatment with YtnP-lactonase decreased the relative mRNA levels of genes involved in the production of elastase (lasB approximately 2-fold), alginate (algK approximately 2.2-fold), pyocyanin (phzM approximately 3.5-fold), pyoverdin (pvdS approximately 2-fold), and rhamnolipid (rhlC approximately 4-fold). These results suggest that YtnP lactonase exerts an antivirulence effect at the transcription level. Conclusion: YtnP lactonase, a quorum quenching (QQ) enzyme, has the potential to be used as an innovative enzyme-based antivirulence therapeutic to combat infections caused by P. aeruginosa. PB - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade C3 - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia T1 - A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression EP - 121 SP - 121 UR - https://hdl.handle.net/21.15107/rcub_intor_803 ER -
@conference{ author = "Ćurčić, Jovana and Jakovljević, Stefan and Novović, Katarina and Vasiljević, Zorica and Kojić, Milan and Jovčić, Branko and Malešević, Milka", year = "2023", abstract = "Introduction: Quorum quenching (QQ) isthe enzymatic degradation of cell-to-cellsignaling molecules. In this study, the potential of the novel YtnP lactonase, the quorum quenching enzyme derived from S. maltophilia, to reduce P. aeruginosa quorum sensing and virulence factor gene expression was investigated. Methods: MMA83 culture (adjusted to 1.5x105 CFU/ml) was treated with recombinant YtnP lactonase (final concentration 50 μg/ml) at 37°C for 12 hours under aeration. RNA isolation of the treated and untreated MMA83 culture was performed using the RNeasy Mini Kit (Qiagen, Germany) according to the protocol. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR), was used to analyze the effect ofYtnP lactonase on the relative mRNA levels of the LasI/LasR, RhiI/RhiR, and PQS signaling network genes of P. aeruginosa MMA83 and virulence factor genes. The rpsL was used as an endogenous control to normalize obtained data following the 2-ΔΔCt method. Results: The QS genes belonging to three QS networks – LasI/LasR, RhiI/RhiR, and PQS of P. aeruginosa MMA83 treated with YtnP lactonase were significantly downregulated. The RT -qPCR results show that treatment with YtnP-lactonase decreased the relative mRNA levels of genes involved in the production of elastase (lasB approximately 2-fold), alginate (algK approximately 2.2-fold), pyocyanin (phzM approximately 3.5-fold), pyoverdin (pvdS approximately 2-fold), and rhamnolipid (rhlC approximately 4-fold). These results suggest that YtnP lactonase exerts an antivirulence effect at the transcription level. Conclusion: YtnP lactonase, a quorum quenching (QQ) enzyme, has the potential to be used as an innovative enzyme-based antivirulence therapeutic to combat infections caused by P. aeruginosa.", publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade", journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia", title = "A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression", pages = "121-121", url = "https://hdl.handle.net/21.15107/rcub_intor_803" }
Ćurčić, J., Jakovljević, S., Novović, K., Vasiljević, Z., Kojić, M., Jovčić, B.,& Malešević, M.. (2023). A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 121-121. https://hdl.handle.net/21.15107/rcub_intor_803
Ćurčić J, Jakovljević S, Novović K, Vasiljević Z, Kojić M, Jovčić B, Malešević M. A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:121-121. https://hdl.handle.net/21.15107/rcub_intor_803 .
Ćurčić, Jovana, Jakovljević, Stefan, Novović, Katarina, Vasiljević, Zorica, Kojić, Milan, Jovčić, Branko, Malešević, Milka, "A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):121-121, https://hdl.handle.net/21.15107/rcub_intor_803 .