de novo pyrimidine synthesis pathway inhibition reduces motility virulence of Pseudomonas aeruginosa despite complementation
Objectives: Finding alternative methods to overcome antibiotic resistance in the medically important bacterium Pseudomonas aeruginosa has become increasingly vital as antibiotic resistance becomes more difficult to overcome. DNA synthesis inhibition is a strategy that has been used to inhibit cancer...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-06-01
|
| Series: | Journal of King Saud University: Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S101836472200221X |
| _version_ | 1817976913561387008 |
|---|---|
| author | Abdurahman A. Niazy Rhodanne Nicole A. Lambarte Hamdan S. Alghamdi |
| author_facet | Abdurahman A. Niazy Rhodanne Nicole A. Lambarte Hamdan S. Alghamdi |
| author_sort | Abdurahman A. Niazy |
| collection | DOAJ |
| description | Objectives: Finding alternative methods to overcome antibiotic resistance in the medically important bacterium Pseudomonas aeruginosa has become increasingly vital as antibiotic resistance becomes more difficult to overcome. DNA synthesis inhibition is a strategy that has been used to inhibit cancer cells from proliferating by using pyrimidine analogues. Gateway site-directed mutagenesis is an effective way to create precise knockouts which creates good models to understand the relationship between biochemical pathways and the physiology of the organism. This article demonstrates the effect of pyrimidine synthesis inhibition on the motility virulence and physiology of P. aeruginosa. Methods: Wild type Pseudomonas aeruginosa (PA01) was genetically modified using gateway cloning technology to inhibit the pyrimidine synthesis pathway through the complete knockout of the pyrE gene. The mutant strain was compared to the wild type PA01 in terms of its twitching, swarming and swimming motility forms and biofilm formation abilities. In addition to that, the production of the bacterial surfactant, rhamnolipids, was also measured. Results and conclusions: The inhibition of the pyrimidine synthesis pathway resulted in significant reduction in motility and biofilm capabilities, despite complementing the mutation with uracil to bypass the mutation. Inhibition of the pyrimidine synthesis pathway is thus an effective way to reduce the virulence of Pseudomonas aeruginosa. Even though the bacterium is able to acquire pyrimidines from its surrounding, it was not sufficient enough to overcome the effects of the mutation. |
| first_indexed | 2024-04-13T22:09:08Z |
| format | Article |
| id | doaj.art-97e19b7607ca44b0979350d2853da160 |
| institution | Directory Open Access Journal |
| issn | 1018-3647 |
| language | English |
| last_indexed | 2024-04-13T22:09:08Z |
| publishDate | 2022-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of King Saud University: Science |
| spelling | doaj.art-97e19b7607ca44b0979350d2853da1602022-12-22T02:27:48ZengElsevierJournal of King Saud University: Science1018-36472022-06-01344102040de novo pyrimidine synthesis pathway inhibition reduces motility virulence of Pseudomonas aeruginosa despite complementationAbdurahman A. Niazy0Rhodanne Nicole A. Lambarte1Hamdan S. Alghamdi2Department of Oral Medicine and Diagnostic Sciences, College of Dentistry, King Saud University, Riyadh, Saudi Arabia; Molecular and Cell Biology Laboratory, Prince Naif bin AbdulAziz Health Research Center, King Saud University, Riyadh, Saudi Arabia; Corresponding author at: Department of Oral Medicine and Diagnostic Sciences, College of Dentistry, King Saud University, Riyadh, Saudi Arabia.Molecular and Cell Biology Laboratory, Prince Naif bin AbdulAziz Health Research Center, King Saud University, Riyadh, Saudi ArabiaDepartment of Periodontics and Community Dentistry, College of Dentistry, King Saud University, Riyadh, Saudi ArabiaObjectives: Finding alternative methods to overcome antibiotic resistance in the medically important bacterium Pseudomonas aeruginosa has become increasingly vital as antibiotic resistance becomes more difficult to overcome. DNA synthesis inhibition is a strategy that has been used to inhibit cancer cells from proliferating by using pyrimidine analogues. Gateway site-directed mutagenesis is an effective way to create precise knockouts which creates good models to understand the relationship between biochemical pathways and the physiology of the organism. This article demonstrates the effect of pyrimidine synthesis inhibition on the motility virulence and physiology of P. aeruginosa. Methods: Wild type Pseudomonas aeruginosa (PA01) was genetically modified using gateway cloning technology to inhibit the pyrimidine synthesis pathway through the complete knockout of the pyrE gene. The mutant strain was compared to the wild type PA01 in terms of its twitching, swarming and swimming motility forms and biofilm formation abilities. In addition to that, the production of the bacterial surfactant, rhamnolipids, was also measured. Results and conclusions: The inhibition of the pyrimidine synthesis pathway resulted in significant reduction in motility and biofilm capabilities, despite complementing the mutation with uracil to bypass the mutation. Inhibition of the pyrimidine synthesis pathway is thus an effective way to reduce the virulence of Pseudomonas aeruginosa. Even though the bacterium is able to acquire pyrimidines from its surrounding, it was not sufficient enough to overcome the effects of the mutation.http://www.sciencedirect.com/science/article/pii/S101836472200221XPseudomonas aeruginosapyrimidine synthesis pathwaymotilityvirulence factorsbiofilm formationpyrE |
| spellingShingle | Abdurahman A. Niazy Rhodanne Nicole A. Lambarte Hamdan S. Alghamdi de novo pyrimidine synthesis pathway inhibition reduces motility virulence of Pseudomonas aeruginosa despite complementation Journal of King Saud University: Science Pseudomonas aeruginosa pyrimidine synthesis pathway motility virulence factors biofilm formation pyrE |
| title | de novo pyrimidine synthesis pathway inhibition reduces motility virulence of Pseudomonas aeruginosa despite complementation |
| title_full | de novo pyrimidine synthesis pathway inhibition reduces motility virulence of Pseudomonas aeruginosa despite complementation |
| title_fullStr | de novo pyrimidine synthesis pathway inhibition reduces motility virulence of Pseudomonas aeruginosa despite complementation |
| title_full_unstemmed | de novo pyrimidine synthesis pathway inhibition reduces motility virulence of Pseudomonas aeruginosa despite complementation |
| title_short | de novo pyrimidine synthesis pathway inhibition reduces motility virulence of Pseudomonas aeruginosa despite complementation |
| title_sort | de novo pyrimidine synthesis pathway inhibition reduces motility virulence of pseudomonas aeruginosa despite complementation |
| topic | Pseudomonas aeruginosa pyrimidine synthesis pathway motility virulence factors biofilm formation pyrE |
| url | http://www.sciencedirect.com/science/article/pii/S101836472200221X |
| work_keys_str_mv | AT abdurahmananiazy denovopyrimidinesynthesispathwayinhibitionreducesmotilityvirulenceofpseudomonasaeruginosadespitecomplementation AT rhodannenicolealambarte denovopyrimidinesynthesispathwayinhibitionreducesmotilityvirulenceofpseudomonasaeruginosadespitecomplementation AT hamdansalghamdi denovopyrimidinesynthesispathwayinhibitionreducesmotilityvirulenceofpseudomonasaeruginosadespitecomplementation |