Pathogenicity and genomic features of vapN-harboring Rhodococcus equi isolated from human patients

Suzuki, Yasunori; Kubota, Hiroaki; Madarame, Hiroo; Takase, Fumiaki; Takahashi, Kei; Sasaki, Yukako; Kakuda, Tsutomu; Takaı̈, Shinji

doi:10.1016/j.ijmm.2021.151519

Cited by 18 publications

(16 citation statements)

References 39 publications

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“…Genes related to secretory protein( pknG) , protease ( zmp1 ), Immune evasion ( pgi ), Cell surface components ( mymA ), anaerobic respiration ( narH , narG ), amino acid and purine metabolism ( lysA ), regulation ( phoR ) and iron uptake (mbtI , fadE14 , fagB , fagC ) were found in all the studied strains. pknG gene was considered to play a core role in Rhocs, a newly discovered heavy oxygen home static system to regulate biofilm growth and host persistence in mycobacteria ( Suzuki et al., 2021 ). NarG gene encodes nitrate reductase G, which participate in respiration under anaerobic conditions.…”

Section: Discussionmentioning

confidence: 99%

“…So far, three types of plasmids have been identified: type-A, B and N, carrying specific VAPs highly related to different humans and animal hosts ( MacArthur et al., 2017 ; Vázquez Boland and Meijer, 2019 ). In humans, most R. equi isolates carry type-B plasmids, while type-A and type-N plasmids are relatively few ( Valero-Rello et al., 2015 ; Suzuki et al., 2021 ). In previous studies, strains carrying type-A plasmids were regarded as high toxic, strains carrying type-B plasmids as medium toxic, type-A and type-B plasmids negative strains as non-toxic ( Jain et al., 2003 ).…”

Section: Introductionmentioning

confidence: 99%

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Genomic Characteristics and Pan-Genome Analysis of Rhodococcus equi

Song

Huang

et al. 2022

Front. Cell. Infect. Microbiol.

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Rhodococcus equi is a zoonotic pathogen that can cause fatal disease in patients who are immunocompromised. At present, the epidemiology and pathogenic mechanisms of R. equi infection are not clear. This study characterized the genomes of 53 R. equi strains from different sources. Pan-genome analysis showed that all R. equi strains contained 11481 pan genes, including 3690 core genes and 602 ~ 1079 accessory genes. Functional annotation of pan genome focused on the genes related to basic lifestyle, such as the storage and expression of metabolic and genetic information. Phylogenetic analysis based on pan-genome showed that the R. equi strains were clustered into six clades, which was not directly related to the isolation location and host source. Also, a total of 84 virulence genes were predicted in 53 R. equi strains. These virulence factors can be divided into 20 categories related to substance metabolism, secreted protein and immune escape. Meanwhile, six antibiotic resistance genes (RbpA, tetA (33), erm (46), sul1, qacEdelta 1 and aadA9) were detected, and all strains carried RbpA related to rifamycin resistance. In addition, 28 plasmids were found in the 53 R. equi strains, belonging to Type-A (n = 14), Type-B (n = 8) and Type-N (n = 6), respectively. The genetic structures of the same type of plasmid were highly similar. In conclusion, R. equi strains show different genomic characteristics, virulence-related genes, potential drug resistance and virulence plasmid structures, which may be conducive to the evolution of its pathogenesis.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Genomic Characteristics and Pan-Genome Analysis of Rhodococcus equi

Song

Huang

et al. 2022

Front. Cell. Infect. Microbiol.

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“…The two DNA fragments were then ligated with the NEBuilder HiFi DNA Assembly Master Mix (New England Biolabs) and directly transformed into NEB 10‐beta to generate pDelta::pVAPA39534_kanR_39535 (pYSK2). pYSKR2 was transformed into U19 or ATCC33701, as described previously 3,21 . In brief, 1 μg pYSKR2 was electroporated into U19 or ATCC33701, and transformants (single crossovers) were selected on LB agar containing 50 μg/mL apramycin and 50 μg/mL kanamycin (FUJIFILM Wako Pure Chemical Corporation), generating YSKR1 and YSKR2.…”

Section: Materials and Methodsmentioning

confidence: 99%

“…pYSKR2 was transformed into U19 or ATCC33701, as described previously. 3,21 In brief, 1 μg pYSKR2 was electroporated into U19 or ATCC33701, and transformants (single crossovers) were selected on LB agar containing 50 μg/mL apramycin and 50 μg/mL kanamycin (FUJIFILM Wako Pure Chemical Corporation), generating YSKR1 and YSKR2.…”

Section: Plasmid Construction and Transformation For R Equimentioning

confidence: 99%

“…1 In addition, the bacterium can be frequently isolated from soil and from purulent lesions of various animals, including humans. [2][3][4][5][6][7] The pathogenicity of R. equi is attributed to its intracellular survival within macrophages, and this property is important for the function of virulence-associated proteins (Vaps) encoded on the virulence plasmid. 8 The major function of Vaps is to generate a pH-neutral, and hence growth-promoting, intracellular niche by preventing the incorporation of the proton-pumping vacuolar ATPase complex into the membrane of phagosomes and the subsequent transfer of the complex to lysosomes.…”

Section: Introductionmentioning

confidence: 99%

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Rhodococcus equiU19 strain harbors a nonmobilizable virulence plasmid

Suzuki

Takaı̈

Kubota

et al. 2022

Microbiology and Immunology

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Rhodococcus equiis the causative agent of pyogenic pneumonia in foals, and a virulence‐associated protein A (VapA) encoded on the pVAPA virulence plasmid is important for its pathogenicity. In this study, we analyzed the virulence of R. equi strain U19, originally isolated in the Netherlands in 1997 and the genetic characteristics of the pVAPA_U19 plasmid. U19 expressed VapA that was regulated by temperature and pH and underwent significant intracellular proliferation in macrophages. The restriction fragment length polymorphism of pVAPA_U19 digested with EcoRI was similar to that of pREAT701 (85 kb Type I) harbored by R. equi ATCC33701, although the band pattern at 10–20 kb differed. Whole‐genome sequencing showed that pVAPA_U19 was 51,684 bp in length and that the vapA pathogenicity island region and the replication/participation were almost identical to those in pREAT701. By contrast, the open reading frames (ORF26–ORF45) genes of pREAT701 (approximately 29,000 bp) were absent from pVAPA_U19. In this lacking region, mobility (MOB) genes, such as relaxase, which allow conjugative DNA processing, and the mating pair formation (MPF) genes, which are a form of the Type IV secretion system and provide the mating channel, were present. Coculture between U19 and five different recipient strains (two plasmid‐cured strains and three cryptic plasmid‐harboring strains) demonstrated that pVAPA_U19 could not support conjugation. Therefore, pVAPA_U19 does not differ significantly from the previously reported pVAPA in terms of virulence and plasmid replication and maintenance but is a nonmobilizable plasmid unable to cause conjugation because of the absence of genes related to MOB and MPF.

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