Transcriptome analysis of Pseudomonas aeruginosa after MhqO dioxygenase treatment
Authors
Đokić, AnđelaMorić, Ivana
Šenerović, Lidija
Đokić, Lidija
Contributors
Morić, IvanaĐorđević, Valentina
Conference object (Published version)
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© 2023 Institute of Molecular Genetics and Genetic Engineering, University of Belgrade
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Show full item recordAbstract
Pseudomonas aeruginosa is an opportunistic pathogen that can cause severe chronic
infections due to its exceptional ability to form a biofilm. Regulation of biofilm formation
is very sophisticated and involves multiple bacterial systems and regulatory pathways.
We found an enzyme MhqO dioxygenase from Bacillus paralicheniformis ZP1, which
was effective in the inhibition of biofilm formation and disruption of mature biofilm
of P. aeruginosa. Our results suggest that MhqO exerts its effect at the adhesion level,
preventing cells from attaching to the surface. We have also shown that the enzyme
stimulates the rhamnolipids synthesis.
To elucidate the mechanism of enzyme action, we analyzed the transcriptome of the P.
aeruginosa PAO1 strain treated with MhqO. Since cell adhesion occurs at the beginning
of the stationary phase growth, the PAO1 strain was treated with MhqO for four hours,
followed by total RNA isolation and cDNA synthesis. Transcriptome sequencing was
performed by... Illumina NovaSeq 6000 and data were analyzed by Novogene Bioinformatics
Technology Co., Ltd. (Beijing, China).
Obtained data showed that 122 genes were up-regulated, 41 genes were down-regulated,
and the expression of 5947 genes was not changed. Five genes whose expression was
altered are directly related to biofilm formation. MhqO increased the expression of the
RsmA post-transcriptional regulator in P. aeruginosa. Transcriptome data revealed that pili
IV biosynthesis genes were up-regulated, which is in accordance with literature data that
RsmA positively regulates these genes. The inhibition of cells’ attachment to the surface
could be explained by these results. In addition, RsmA positively regulates rhamnolipid
production but negatively regulates biofilm matrix synthesis, which was supported by
expression levels in the sequenced transcriptome.
Data obtained from transcriptome analysis suggest that P. aeruginosa treated with MhqO
dioxygenase should be more sensitive to oxidative and osmotic stress, as well as to
beta-lactam antibiotics. Our further investigations should confirm these effects at the
phenotypic level as well.
Keywords:
transcriptome / MhqO / RsmA / Pseudomonas aeruginosaSource:
4th Belgrade Bioinformatics Conference, 2023, 4, 102-102Publisher:
- Belgrade : Institute of molecular genetics and genetic engineering
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)
Note:
- Book of abstract: 4th Belgrade Bioinformatics Conference, June 19-23, 2023
Institution/Community
Institut za molekularnu genetiku i genetičko inženjerstvoTY - CONF AU - Đokić, Anđela AU - Morić, Ivana AU - Šenerović, Lidija AU - Đokić, Lidija PY - 2023 UR - https://belbi.bg.ac.rs/ UR - https://imagine.imgge.bg.ac.rs/handle/123456789/2047 AB - Pseudomonas aeruginosa is an opportunistic pathogen that can cause severe chronic infections due to its exceptional ability to form a biofilm. Regulation of biofilm formation is very sophisticated and involves multiple bacterial systems and regulatory pathways. We found an enzyme MhqO dioxygenase from Bacillus paralicheniformis ZP1, which was effective in the inhibition of biofilm formation and disruption of mature biofilm of P. aeruginosa. Our results suggest that MhqO exerts its effect at the adhesion level, preventing cells from attaching to the surface. We have also shown that the enzyme stimulates the rhamnolipids synthesis. To elucidate the mechanism of enzyme action, we analyzed the transcriptome of the P. aeruginosa PAO1 strain treated with MhqO. Since cell adhesion occurs at the beginning of the stationary phase growth, the PAO1 strain was treated with MhqO for four hours, followed by total RNA isolation and cDNA synthesis. Transcriptome sequencing was performed by Illumina NovaSeq 6000 and data were analyzed by Novogene Bioinformatics Technology Co., Ltd. (Beijing, China). Obtained data showed that 122 genes were up-regulated, 41 genes were down-regulated, and the expression of 5947 genes was not changed. Five genes whose expression was altered are directly related to biofilm formation. MhqO increased the expression of the RsmA post-transcriptional regulator in P. aeruginosa. Transcriptome data revealed that pili IV biosynthesis genes were up-regulated, which is in accordance with literature data that RsmA positively regulates these genes. The inhibition of cells’ attachment to the surface could be explained by these results. In addition, RsmA positively regulates rhamnolipid production but negatively regulates biofilm matrix synthesis, which was supported by expression levels in the sequenced transcriptome. Data obtained from transcriptome analysis suggest that P. aeruginosa treated with MhqO dioxygenase should be more sensitive to oxidative and osmotic stress, as well as to beta-lactam antibiotics. Our further investigations should confirm these effects at the phenotypic level as well. PB - Belgrade : Institute of molecular genetics and genetic engineering C3 - 4th Belgrade Bioinformatics Conference T1 - Transcriptome analysis of Pseudomonas aeruginosa after MhqO dioxygenase treatment EP - 102 SP - 102 VL - 4 UR - https://hdl.handle.net/21.15107/rcub_imagine_2047 ER -
@conference{ author = "Đokić, Anđela and Morić, Ivana and Šenerović, Lidija and Đokić, Lidija", year = "2023", abstract = "Pseudomonas aeruginosa is an opportunistic pathogen that can cause severe chronic infections due to its exceptional ability to form a biofilm. Regulation of biofilm formation is very sophisticated and involves multiple bacterial systems and regulatory pathways. We found an enzyme MhqO dioxygenase from Bacillus paralicheniformis ZP1, which was effective in the inhibition of biofilm formation and disruption of mature biofilm of P. aeruginosa. Our results suggest that MhqO exerts its effect at the adhesion level, preventing cells from attaching to the surface. We have also shown that the enzyme stimulates the rhamnolipids synthesis. To elucidate the mechanism of enzyme action, we analyzed the transcriptome of the P. aeruginosa PAO1 strain treated with MhqO. Since cell adhesion occurs at the beginning of the stationary phase growth, the PAO1 strain was treated with MhqO for four hours, followed by total RNA isolation and cDNA synthesis. Transcriptome sequencing was performed by Illumina NovaSeq 6000 and data were analyzed by Novogene Bioinformatics Technology Co., Ltd. (Beijing, China). Obtained data showed that 122 genes were up-regulated, 41 genes were down-regulated, and the expression of 5947 genes was not changed. Five genes whose expression was altered are directly related to biofilm formation. MhqO increased the expression of the RsmA post-transcriptional regulator in P. aeruginosa. Transcriptome data revealed that pili IV biosynthesis genes were up-regulated, which is in accordance with literature data that RsmA positively regulates these genes. The inhibition of cells’ attachment to the surface could be explained by these results. In addition, RsmA positively regulates rhamnolipid production but negatively regulates biofilm matrix synthesis, which was supported by expression levels in the sequenced transcriptome. Data obtained from transcriptome analysis suggest that P. aeruginosa treated with MhqO dioxygenase should be more sensitive to oxidative and osmotic stress, as well as to beta-lactam antibiotics. Our further investigations should confirm these effects at the phenotypic level as well.", publisher = "Belgrade : Institute of molecular genetics and genetic engineering", journal = "4th Belgrade Bioinformatics Conference", title = "Transcriptome analysis of Pseudomonas aeruginosa after MhqO dioxygenase treatment", pages = "102-102", volume = "4", url = "https://hdl.handle.net/21.15107/rcub_imagine_2047" }
Đokić, A., Morić, I., Šenerović, L.,& Đokić, L.. (2023). Transcriptome analysis of Pseudomonas aeruginosa after MhqO dioxygenase treatment. in 4th Belgrade Bioinformatics Conference Belgrade : Institute of molecular genetics and genetic engineering., 4, 102-102. https://hdl.handle.net/21.15107/rcub_imagine_2047
Đokić A, Morić I, Šenerović L, Đokić L. Transcriptome analysis of Pseudomonas aeruginosa after MhqO dioxygenase treatment. in 4th Belgrade Bioinformatics Conference. 2023;4:102-102. https://hdl.handle.net/21.15107/rcub_imagine_2047 .
Đokić, Anđela, Morić, Ivana, Šenerović, Lidija, Đokić, Lidija, "Transcriptome analysis of Pseudomonas aeruginosa after MhqO dioxygenase treatment" in 4th Belgrade Bioinformatics Conference, 4 (2023):102-102, https://hdl.handle.net/21.15107/rcub_imagine_2047 .