A novel mutation of walK confers vancomycin-intermediate resistance in methicillin-susceptible Staphylococcus aureus
With the treatment failure by vancomycin and poor clinical outcomes, the emergence and spread of vancomycin intermediate-resistant Staphylococcus aureus (VISA) has raised more concerns in recent years. While most VISA strains are isolated from methicillin-resistant S. aureus (MRSA), the mechanism un...
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| Published in | International journal of medical microbiology Vol. 311; no. 2; p. 151473 |
|---|---|
| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
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Germany
Elsevier GmbH
01.02.2021
Elsevier |
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| Abstract | With the treatment failure by vancomycin and poor clinical outcomes, the emergence and spread of vancomycin intermediate-resistant Staphylococcus aureus (VISA) has raised more concerns in recent years. While most VISA strains are isolated from methicillin-resistant S. aureus (MRSA), the mechanism underlying the generation of VISA from methicillin-susceptible S. aureus (MSSA) is still largely unknown. Here, we identified a total of 10 mutations in 9 genes through comparative genome analysis from laboratory-derived VISA strain. We verified the role of a novel mutation of WalK (I237T) and our results further indicated that the introduction of WalK (I237T) by allelic replacement can confer vancomycin resistance in MSSA with common VISA characteristics, including thickened cell walls, reduced autolysis, and attenuated virulence. Consistent with these phenotypes, real-time quantitative reverse transcription-PCR revealed the altered expression of several genes associated with cell wall metabolism and virulence control. In addition, electrophoretic mobility shift assay indicated that WalR can directly bind to the promoter regions of oatA, sle1, and mgt, fluorescence-based promoter activity and β-galactosidase assays revealed WalK (I237T) can alter promoter activities of oatA, mgt, and sle1, thus regulating genes expression. These findings broaden our understanding of the regulatory network by WalKR system and decipher the molecular mechanisms of developmental VISA resistance in MSSA with point mutations. |
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| AbstractList | With the treatment failure by vancomycin and poor clinical outcomes, the emergence and spread of vancomycin intermediate-resistant Staphylococcus aureus (VISA) has raised more concerns in recent years. While most VISA strains are isolated from methicillin-resistant S. aureus (MRSA), the mechanism underlying the generation of VISA from methicillin-susceptible S. aureus (MSSA) is still largely unknown. Here, we identified a total of 10 mutations in 9 genes through comparative genome analysis from laboratory-derived VISA strain. We verified the role of a novel mutation of WalK (I237T) and our results further indicated that the introduction of WalK (I237T) by allelic replacement can confer vancomycin resistance in MSSA with common VISA characteristics, including thickened cell walls, reduced autolysis, and attenuated virulence. Consistent with these phenotypes, real-time quantitative reverse transcription-PCR revealed the altered expression of several genes associated with cell wall metabolism and virulence control. In addition, electrophoretic mobility shift assay indicated that WalR can directly bind to the promoter regions of oatA, sle1, and mgt, fluorescence-based promoter activity and β-galactosidase assays revealed WalK (I237T) can alter promoter activities of oatA, mgt, and sle1, thus regulating genes expression. These findings broaden our understanding of the regulatory network by WalKR system and decipher the molecular mechanisms of developmental VISA resistance in MSSA with point mutations. |
| ArticleNumber | 151473 |
| Author | Zhu, Jiade Shu, Xueqin Liu, Banghui Sun, Baolin |
| Author_xml | – sequence: 1 givenname: Jiade surname: Zhu fullname: Zhu, Jiade – sequence: 2 givenname: Banghui surname: Liu fullname: Liu, Banghui – sequence: 3 givenname: Xueqin surname: Shu fullname: Shu, Xueqin – sequence: 4 givenname: Baolin surname: Sun fullname: Sun, Baolin email: sunb@ustc.edu.cn |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/33445057$$D View this record in MEDLINE/PubMed |
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| CitedBy_id | crossref_primary_10_3103_S0891416824700071 crossref_primary_10_1186_s12934_023_02018_0 crossref_primary_10_1128_spectrum_03596_22 crossref_primary_10_1128_jb_00418_23 crossref_primary_10_2147_IDR_S411860 crossref_primary_10_3389_fmicb_2021_696730 crossref_primary_10_36488_cmac_2023_2_187_201 crossref_primary_10_1128_spectrum_02567_22 crossref_primary_10_3389_fmicb_2022_820089 crossref_primary_10_1134_S1068162024030087 crossref_primary_10_1186_s12866_021_02298_9 |
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| Keywords | Vancomycin-intermediate Staphylococcus aureus Vancomycin resistance WalKR Point mutation |
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| SubjectTerms | Anti-Bacterial Agents - pharmacology Comparative Genomic Hybridization Genes, Bacterial Methicillin - pharmacology Microbial Sensitivity Tests Mutation Point mutation Staphylococcus aureus - drug effects Staphylococcus aureus - genetics Vancomycin - pharmacology Vancomycin resistance Vancomycin Resistance - genetics Vancomycin-intermediate Staphylococcus aureus WalKR |
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| Title | A novel mutation of walK confers vancomycin-intermediate resistance in methicillin-susceptible Staphylococcus aureus |
| URI | https://dx.doi.org/10.1016/j.ijmm.2021.151473 https://www.ncbi.nlm.nih.gov/pubmed/33445057 https://search.proquest.com/docview/2478585069 https://doaj.org/article/da5203fcaf384ae4afd97e46f8d40bb6 |
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