The nitric oxide synthase gene negatively regulates biofilm formation in Staphylococcus epidermidis
Staphylococcus epidermidis (S. epidermidis) is a clinically important conditioned pathogen that can cause a troublesome chronic implant-related infection once a biofilm is formed. The nitric oxide synthase (NOS) gene, which is responsible for endogenous nitric oxide synthesis, has already been found...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2022.1015859/full |
| _version_ | 1811192799861669888 |
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| author | Jiaxue Wang Jiaxue Wang Jiaxue Wang Jiaxue Wang Lulin Rao Zhuoan Huang Lili Ma Tian Yang Zhongqi Yu Aihua Sun Yumei Ge Yumei Ge Yumei Ge Yumei Ge |
| author_facet | Jiaxue Wang Jiaxue Wang Jiaxue Wang Jiaxue Wang Lulin Rao Zhuoan Huang Lili Ma Tian Yang Zhongqi Yu Aihua Sun Yumei Ge Yumei Ge Yumei Ge Yumei Ge |
| author_sort | Jiaxue Wang |
| collection | DOAJ |
| description | Staphylococcus epidermidis (S. epidermidis) is a clinically important conditioned pathogen that can cause a troublesome chronic implant-related infection once a biofilm is formed. The nitric oxide synthase (NOS) gene, which is responsible for endogenous nitric oxide synthesis, has already been found in the genome of S. epidermidis; however, the specific mechanisms associated with the effects of NOS on S. epidermidis pathogenicity are still unknown. The purpose of the current study was to investigate whether the NOS gene has an impact on biofilm formation in S. epidermidis. Bioinformatics analysis of the NOS gene was performed, and homologous recombination was subsequently employed to delete this gene. The effects of the NOS gene on biofilm formation of S. epidermidis and its underlying mechanisms were analyzed by bacterial growth assays, biofilm semiquantitative determination, Triton X-100-induced autolysis assays, and bacterial biofilm dispersal assays. Additionally, the transcription levels of fbe, aap, icaA, icaR and sigB, which are related to biofilm formation, were further investigated by qRT-PCR following NOS deletion. Phylogenetic analysis revealed that the NOS gene was conserved between bacterial species originating from different genera. The NOS deletion strain of S. epidermidis 1457 and its counterpart were successfully constructed. Disruption of the NOS gene resulted in significantly enhanced biofilm formation, slightly retarded bacterial growth, a markedly decreased autolysis rate, and drastically weakened bacterial biofilm dispersal. Our data showed that the fbe, aap and icaA genes were significantly upregulated, while the icaR and sigB genes were significantly downregulated, compared with the wild strain. Therefore, these data strongly suggested that the NOS gene can negatively regulate biofilm formation in S. epidermidis by affecting biofilm aggregation and dispersal. |
| first_indexed | 2024-04-11T23:57:34Z |
| format | Article |
| id | doaj.art-7e224fa9c4ba432bac86b7971bb0c9cb |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-04-11T23:57:34Z |
| publishDate | 2022-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-7e224fa9c4ba432bac86b7971bb0c9cb2022-12-22T03:56:19ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882022-11-011210.3389/fcimb.2022.10158591015859The nitric oxide synthase gene negatively regulates biofilm formation in Staphylococcus epidermidisJiaxue Wang0Jiaxue Wang1Jiaxue Wang2Jiaxue Wang3Lulin Rao4Zhuoan Huang5Lili Ma6Tian Yang7Zhongqi Yu8Aihua Sun9Yumei Ge10Yumei Ge11Yumei Ge12Yumei Ge13School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, Zhejiang, ChinaDepartment of Clinical Laboratory, Laboratory Medicine Center, Zhejiang Provincial People’s Hospital (Affiliated People’s Hospital, Hangzhou Medical College), Hangzhou, Zhejiang, ChinaKey Laboratory of Biomarkers and In Vitro Diagnosis Translation of Zhejiang province, Hangzhou, Zhejiang, ChinaInstitute of Clinical Microbiology, Hangzhou Medical College, Hangzhou, Zhejiang, ChinaSchool of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, Zhejiang, ChinaSchool of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, Zhejiang, ChinaSchool of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, Zhejiang, ChinaSchool of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, Zhejiang, ChinaSchool of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, Zhejiang, ChinaDepartment of basic medicine, Hangzhou Medical College, Hangzhou, Zhejiang, ChinaDepartment of Clinical Laboratory, Laboratory Medicine Center, Zhejiang Provincial People’s Hospital (Affiliated People’s Hospital, Hangzhou Medical College), Hangzhou, Zhejiang, ChinaKey Laboratory of Biomarkers and In Vitro Diagnosis Translation of Zhejiang province, Hangzhou, Zhejiang, ChinaInstitute of Clinical Microbiology, Hangzhou Medical College, Hangzhou, Zhejiang, ChinaDepartment of basic medicine, Hangzhou Medical College, Hangzhou, Zhejiang, ChinaStaphylococcus epidermidis (S. epidermidis) is a clinically important conditioned pathogen that can cause a troublesome chronic implant-related infection once a biofilm is formed. The nitric oxide synthase (NOS) gene, which is responsible for endogenous nitric oxide synthesis, has already been found in the genome of S. epidermidis; however, the specific mechanisms associated with the effects of NOS on S. epidermidis pathogenicity are still unknown. The purpose of the current study was to investigate whether the NOS gene has an impact on biofilm formation in S. epidermidis. Bioinformatics analysis of the NOS gene was performed, and homologous recombination was subsequently employed to delete this gene. The effects of the NOS gene on biofilm formation of S. epidermidis and its underlying mechanisms were analyzed by bacterial growth assays, biofilm semiquantitative determination, Triton X-100-induced autolysis assays, and bacterial biofilm dispersal assays. Additionally, the transcription levels of fbe, aap, icaA, icaR and sigB, which are related to biofilm formation, were further investigated by qRT-PCR following NOS deletion. Phylogenetic analysis revealed that the NOS gene was conserved between bacterial species originating from different genera. The NOS deletion strain of S. epidermidis 1457 and its counterpart were successfully constructed. Disruption of the NOS gene resulted in significantly enhanced biofilm formation, slightly retarded bacterial growth, a markedly decreased autolysis rate, and drastically weakened bacterial biofilm dispersal. Our data showed that the fbe, aap and icaA genes were significantly upregulated, while the icaR and sigB genes were significantly downregulated, compared with the wild strain. Therefore, these data strongly suggested that the NOS gene can negatively regulate biofilm formation in S. epidermidis by affecting biofilm aggregation and dispersal.https://www.frontiersin.org/articles/10.3389/fcimb.2022.1015859/fullnitric oxide synthase geneS. epidermidisnitric oxidebiofilmchronic infection |
| spellingShingle | Jiaxue Wang Jiaxue Wang Jiaxue Wang Jiaxue Wang Lulin Rao Zhuoan Huang Lili Ma Tian Yang Zhongqi Yu Aihua Sun Yumei Ge Yumei Ge Yumei Ge Yumei Ge The nitric oxide synthase gene negatively regulates biofilm formation in Staphylococcus epidermidis Frontiers in Cellular and Infection Microbiology nitric oxide synthase gene S. epidermidis nitric oxide biofilm chronic infection |
| title | The nitric oxide synthase gene negatively regulates biofilm formation in Staphylococcus epidermidis |
| title_full | The nitric oxide synthase gene negatively regulates biofilm formation in Staphylococcus epidermidis |
| title_fullStr | The nitric oxide synthase gene negatively regulates biofilm formation in Staphylococcus epidermidis |
| title_full_unstemmed | The nitric oxide synthase gene negatively regulates biofilm formation in Staphylococcus epidermidis |
| title_short | The nitric oxide synthase gene negatively regulates biofilm formation in Staphylococcus epidermidis |
| title_sort | nitric oxide synthase gene negatively regulates biofilm formation in staphylococcus epidermidis |
| topic | nitric oxide synthase gene S. epidermidis nitric oxide biofilm chronic infection |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2022.1015859/full |
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