Dataset Information

Genome Sequence of a Highly Virulent pvl-positive Vancomycin-intermediate-resistant Staphylococcus aureus Sequence Type 30.

ABSTRACT: Background:Staphylococcus aureus isolates expressing the Panton-Valentine Leukocidin (PVL) have been related to a wide range of diseases. Recently, pvl-positive community-associated methicillin-resistant S. aureus belonging to USA1100 (ST30/CC30/SCCmec IV) lineage has emerged in Brazilian hospitals. Objective:The aim of this work was to sequence the genome of a pvl-positive USA1100 Vancomycin-Intermediate-Resistant S. aureus (VISA) isolate from Rio de Janeiro, Brazil. Methods:The 13420 genome was sequenced using the HiSeq 2500 platform. The draft genome, plasmids annotation, and genome analysis were performed using RAST. Comparison of the relative pvl gene expression of six S. aureus isolates was performed by qRT-PCR. Results:The isolate presented the ?PVL phage codifying for the H2b PVL protein isoform, and another prophage carrying a PVL variant named lukF and lukS-PV.2. The 13420 genome presented a high number of virulence determinants, such as genes codifying for serine-protease proteins, enterotoxins (egc), the immune evasion cluster (IEC), adhesion proteins, spermine/spermidine acetyltransferase gene (blt), superantigen-like proteins, as well as the ica operon. Point mutations at vraS, tcaA, and tcaB genes were detected. Moreover, the PVL mRNA relative expression of the 13420 isolate was five times higher than mRNA PVL levels of the USA300/ST8 reference strain. Conclusion:We described for the first time the genome sequence of a VISA isolate harboring two pvl-associated genes and other virulence factors that may improve the USA1100/ST30 lineage fitness and impact its pathogenicity and spreading at Brazilian hospitals.

SUBMITTER: Chamon RC

PROVIDER: S-EPMC7324871 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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Genome Sequence of a Highly Virulent <i>pvl</i>-positive Vancomycin-intermediate-resistant <i>Staphylococcus aureus</i> Sequence Type 30.

Chamon Raiane C RC Marques Lucas M LM Timenetsky Jorge J da Costa Rachid Caio T C CTC Ferreira Rosana B R RBR de Oliveira Tamara L R TLR Glatthardt Thais T de Oliveira Moreira Lilian L Dos Santos Kátia R N KRN

Current genomics 20200201 2

<h4>Background</h4><i>Staphylococcus aureus</i> isolates expressing the Panton-Valentine Leukocidin (PVL) have been related to a wide range of diseases. Recently, <i>pvl</i>-positive community-associated methicillin-resistant <i>S. aureus</i> belonging to USA1100 (ST30/CC30/<i>SCCmec</i> IV) lineage has emerged in Brazilian hospitals.<h4>Objective</h4>The aim of this work was to sequence the genome of a <i>pvl-</i>positive USA1100 Vancomycin-Intermediate-Resistant <i>S. aureus</i> (VISA) isolate ...[more]

PMID: 32655307

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Project description:Vancomycin (VAN)-intermediate Staphylococcus aureus (VISA) and heterogeneous VISA (hVISA) isolates are considered to have emerged from VAN-susceptible S. aureus (VSSA) by spontaneous mutation during VAN exposure. We previously reported that laboratory mutant H14, obtained from VSSA strain Delta IP by exposure to imipenem (IPM), showed overexpression of the vraSR two-component system and a typical hVISA phenotype. In the present study, to elucidate the mechanism of VSSA conversion to hVISA, we further characterized strain H14 by determining its whole-genome sequence, morphology, cell wall synthetic activity, and gene expression. Genome sequencing revealed that H14 harbored a mutated vraS (designated vraS(H14)) that caused an amino acid substitution (S(329)-->L). This mutation is different from the VraS mutation (N(5)-->I) identified in representative clinical hVISA strain Mu3. However, H14 exhibited a phenotype similar to that of Mu3, including heterogeneous resistance to VAN, enhanced cell wall synthetic activity, and vraSR overexpression. Replacement of the vraS gene of DeltaIP with the mutated vraS(H14) gene confirmed that the S(329)-->L substitution was responsible for both the upregulation of vraSR and conversion to the hVISA phenotype. This conversion was also achieved by using the vraS gene of Mu3, which carries a mutation (N(5)-->I), but not with the native vraS gene of strain N315. Finally, we carried out a study to analyze the appearance of hVISA from VSSA by exposure of Delta IP to selective concentrations of VAN and beta-lactam antibiotics. A total of 8 and 5 hVISA isolates were detected among 50 isolates selected with VAN and IPM, respectively. Among the 13 hVISA mutants, mutation in vraSR was detected only in mutant strain H14, suggesting that additional mutational mechanisms can be responsible for evolution to the hVISA phenotype. We conclude that exposure not only to VAN but also to beta-lactams may select for reduced glycopeptide susceptibility in S. aureus.

Dataset Information

Genome Sequence of a Highly Virulent pvl-positive Vancomycin-intermediate-resistant Staphylococcus aureus Sequence Type 30.

Publications

Genome Sequence of a Highly Virulent <i>pvl</i>-positive Vancomycin-intermediate-resistant <i>Staphylococcus aureus</i> Sequence Type 30.

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