Primers used in this study.
<div><p>CwlM, identified as an N-acetylmuramoyl-l-alanine amidase, plays crucial roles in the synthesis and remodeling of peptidoglycan in mycobacteria. This protein also appears to participate in both drug susceptibility and tolerance mechanisms within these organisms. In our study, we...
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2025
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| _version_ | 1852015539104251904 |
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| author | Shufeng Yang (401264) |
| author2 | Yuzhao Ren (22493234) Yu Wu (54264) Xinyang Li (1543561) Xin Liu (43569) Guoying Deng (583775) |
| author2_role | author author author author author |
| author_facet | Shufeng Yang (401264) Yuzhao Ren (22493234) Yu Wu (54264) Xinyang Li (1543561) Xin Liu (43569) Guoying Deng (583775) |
| author_role | author |
| dc.creator.none.fl_str_mv | Shufeng Yang (401264) Yuzhao Ren (22493234) Yu Wu (54264) Xinyang Li (1543561) Xin Liu (43569) Guoying Deng (583775) |
| dc.date.none.fl_str_mv | 2025-10-24T17:39:51Z |
| dc.identifier.none.fl_str_mv | 10.1371/journal.pone.0334937.t001 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/dataset/Primers_used_in_this_study_/30441068 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biophysics Biochemistry Microbiology Cell Biology Genetics Pharmacology Science Policy Infectious Diseases Biological Sciences not elsewhere classified several genes associated protein also appears plays crucial roles employed crispr interference bacterial cell wall increased expression levels towards certain anti mycobacterium smegmatis </ enhanced antibiotic susceptibility div >< p tolerance mechanisms within rif ), pyrazinamide inh ), rifampicin cwlm depletion increased peptidoglycan synthesis appeared smegmatis </ increased permeability tolerance mechanisms elevated expression cwlm depletion type regulator tuberculosis drugs resulting effects pza ), observed tolerance line anti inh might including isoniazid hydrazidase along findings revealed findings indicate emb ), either hydrazidase drug susceptibility deplete cwlm amidase known alanine amidase |
| dc.title.none.fl_str_mv | Primers used in this study. |
| dc.type.none.fl_str_mv | Dataset info:eu-repo/semantics/publishedVersion dataset |
| description | <div><p>CwlM, identified as an N-acetylmuramoyl-l-alanine amidase, plays crucial roles in the synthesis and remodeling of peptidoglycan in mycobacteria. This protein also appears to participate in both drug susceptibility and tolerance mechanisms within these organisms. In our study, we employed CRISPR interference (CRISPRi) to deplete CwlM in <i><i>Mycobacterium smegmatis</i></i> (<i><i>M. smegmatis</i></i>) and examined the resulting effects on the susceptibility of mycobacteria to first-line anti-tuberculosis drugs, including isoniazid (INH), rifampicin (RIF), pyrazinamide (PZA), and ethambutol (EMB), as well as the β-lactams cefoxitin and imipenem. Our findings revealed that CwlM depletion increased the susceptibility of the bacterium to RIF, EMB, cefoxitin, and imipenem, while tolerance was heightened against INH and PZA. The enhanced antibiotic susceptibility can primarily be attributed to increased permeability of the bacterial cell wall. Conversely, the observed tolerance to INH might be ascribed to elevated expression of the amidase known as hydrazidase along with its LuxR-type regulator. Furthermore, several genes associated with peptidoglycan synthesis appeared to correlate with increased expression levels of either hydrazidase or its LuxR-type regulator. Collectively, these findings indicate that CwlM depletion significantly influences the susceptibility of <i><i>M. smegmatis</i></i> towards certain anti-tuberculosis drugs and may be implicated in drug susceptibility and tolerance mechanisms in <i><i>M. smegmatis</i></i>.</p></div> |
| eu_rights_str_mv | openAccess |
| id | Manara_0ec87ae90d1bcfea19c4e99b9564cdfc |
| identifier_str_mv | 10.1371/journal.pone.0334937.t001 |
| network_acronym_str | Manara |
| network_name_str | ManaraRepo |
| oai_identifier_str | oai:figshare.com:article/30441068 |
| publishDate | 2025 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | Primers used in this study.Shufeng Yang (401264)Yuzhao Ren (22493234)Yu Wu (54264)Xinyang Li (1543561)Xin Liu (43569)Guoying Deng (583775)BiophysicsBiochemistryMicrobiologyCell BiologyGeneticsPharmacologyScience PolicyInfectious DiseasesBiological Sciences not elsewhere classifiedseveral genes associatedprotein also appearsplays crucial rolesemployed crispr interferencebacterial cell wallincreased expression levelstowards certain antimycobacterium smegmatis </enhanced antibiotic susceptibilitydiv >< ptolerance mechanisms withinrif ), pyrazinamideinh ), rifampicincwlm depletion increasedpeptidoglycan synthesis appearedsmegmatis </increased permeabilitytolerance mechanismselevated expressioncwlm depletiontype regulatortuberculosis drugsresulting effectspza ),observed toleranceline antiinh mightincluding isoniazidhydrazidase alongfindings revealedfindings indicateemb ),either hydrazidasedrug susceptibilitydeplete cwlmamidase knownalanine amidase<div><p>CwlM, identified as an N-acetylmuramoyl-l-alanine amidase, plays crucial roles in the synthesis and remodeling of peptidoglycan in mycobacteria. This protein also appears to participate in both drug susceptibility and tolerance mechanisms within these organisms. In our study, we employed CRISPR interference (CRISPRi) to deplete CwlM in <i><i>Mycobacterium smegmatis</i></i> (<i><i>M. smegmatis</i></i>) and examined the resulting effects on the susceptibility of mycobacteria to first-line anti-tuberculosis drugs, including isoniazid (INH), rifampicin (RIF), pyrazinamide (PZA), and ethambutol (EMB), as well as the β-lactams cefoxitin and imipenem. Our findings revealed that CwlM depletion increased the susceptibility of the bacterium to RIF, EMB, cefoxitin, and imipenem, while tolerance was heightened against INH and PZA. The enhanced antibiotic susceptibility can primarily be attributed to increased permeability of the bacterial cell wall. Conversely, the observed tolerance to INH might be ascribed to elevated expression of the amidase known as hydrazidase along with its LuxR-type regulator. Furthermore, several genes associated with peptidoglycan synthesis appeared to correlate with increased expression levels of either hydrazidase or its LuxR-type regulator. Collectively, these findings indicate that CwlM depletion significantly influences the susceptibility of <i><i>M. smegmatis</i></i> towards certain anti-tuberculosis drugs and may be implicated in drug susceptibility and tolerance mechanisms in <i><i>M. smegmatis</i></i>.</p></div>2025-10-24T17:39:51ZDatasetinfo:eu-repo/semantics/publishedVersiondataset10.1371/journal.pone.0334937.t001https://figshare.com/articles/dataset/Primers_used_in_this_study_/30441068CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/304410682025-10-24T17:39:51Z |
| spellingShingle | Primers used in this study. Shufeng Yang (401264) Biophysics Biochemistry Microbiology Cell Biology Genetics Pharmacology Science Policy Infectious Diseases Biological Sciences not elsewhere classified several genes associated protein also appears plays crucial roles employed crispr interference bacterial cell wall increased expression levels towards certain anti mycobacterium smegmatis </ enhanced antibiotic susceptibility div >< p tolerance mechanisms within rif ), pyrazinamide inh ), rifampicin cwlm depletion increased peptidoglycan synthesis appeared smegmatis </ increased permeability tolerance mechanisms elevated expression cwlm depletion type regulator tuberculosis drugs resulting effects pza ), observed tolerance line anti inh might including isoniazid hydrazidase along findings revealed findings indicate emb ), either hydrazidase drug susceptibility deplete cwlm amidase known alanine amidase |
| status_str | publishedVersion |
| title | Primers used in this study. |
| title_full | Primers used in this study. |
| title_fullStr | Primers used in this study. |
| title_full_unstemmed | Primers used in this study. |
| title_short | Primers used in this study. |
| title_sort | Primers used in this study. |
| topic | Biophysics Biochemistry Microbiology Cell Biology Genetics Pharmacology Science Policy Infectious Diseases Biological Sciences not elsewhere classified several genes associated protein also appears plays crucial roles employed crispr interference bacterial cell wall increased expression levels towards certain anti mycobacterium smegmatis </ enhanced antibiotic susceptibility div >< p tolerance mechanisms within rif ), pyrazinamide inh ), rifampicin cwlm depletion increased peptidoglycan synthesis appeared smegmatis </ increased permeability tolerance mechanisms elevated expression cwlm depletion type regulator tuberculosis drugs resulting effects pza ), observed tolerance line anti inh might including isoniazid hydrazidase along findings revealed findings indicate emb ), either hydrazidase drug susceptibility deplete cwlm amidase known alanine amidase |