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The VanRS Homologous Two-Component System VnlRS Ab of the Glycopeptide Producer Amycolatopsis balhimycina Activates Transcription of the vanHAX Sc Genes in Streptomyces coelicolor , but not in A. balhimycina

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ABSTRACT

In enterococci and in Streptomyces coelicolor, a glycopeptide nonproducer, the glycopeptide resistance genes vanHAX are colocalized with vanRS. The two-component system (TCS) VanRS activates vanHAX transcription upon sensing the presence of glycopeptides. Amycolatopsis balhimycina, the producer of the vancomycin-like glycopeptide balhimycin, also possesses vanHAXAb genes. The genes for the VanRS-like TCS VnlRSAb, together with the carboxypeptidase gene vanYAb, are part of the balhimycin biosynthetic gene cluster, which is located 2 Mb separate from the vanHAXAb. The deletion of vnlRSAb did not affect glycopeptide resistance or balhimycin production. In the A. balhimycina vnlRAb deletion mutant, the vanHAXAb genes were expressed at the same level as in the wild type, and peptidoglycan (PG) analyses proved the synthesis of resistant PG precursors. Whereas vanHAXAb expression in A. balhimycina does not depend on VnlRAb, a VnlRAb-depending regulation of vanYAb was demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR) and RNA-seq analyses. Although VnlRAb does not regulate the vanHAXAb genes in A. balhimycina, its heterologous expression in the glycopeptide-sensitive S. coelicolor ΔvanRSSc deletion mutant restored glycopeptide resistance. VnlRAb activates the vanHAXSc genes even in the absence of VanS. In addition, expression of vnlRAb increases actinorhodin production and influences morphological differentiation in S. coelicolor.

No MeSH data available.


RT-PCR analyses of vanHAXAb and vanYAb in A. balhimycina WT, A. balhimycina ΔvnlRAb, and in A. balhimycina WT overexpressing vnlRAb (WT [vnlRAb]). RNA was isolated at different time points (15/39/63 hr) from the three strains cultivated in balhimycin production medium R5. sigB: transcription of the housekeeping gene sigB. vanHAXAb and vanYAb: transcription of vanHAXAb and vanYAb. For PCR, genomic DNA (gDNA) was used as positive control.
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f3: RT-PCR analyses of vanHAXAb and vanYAb in A. balhimycina WT, A. balhimycina ΔvnlRAb, and in A. balhimycina WT overexpressing vnlRAb (WT [vnlRAb]). RNA was isolated at different time points (15/39/63 hr) from the three strains cultivated in balhimycin production medium R5. sigB: transcription of the housekeeping gene sigB. vanHAXAb and vanYAb: transcription of vanHAXAb and vanYAb. For PCR, genomic DNA (gDNA) was used as positive control.

Mentions: RT-PCR analyses revealed that a vanHAXAb transcript was detectable in A. balhimycina ΔvnlRAb, confirming that the expression of vanHAXAb is independent of vnlRAb (Fig. 3).


The VanRS Homologous Two-Component System VnlRS Ab of the Glycopeptide Producer Amycolatopsis balhimycina Activates Transcription of the vanHAX Sc Genes in Streptomyces coelicolor , but not in A. balhimycina
RT-PCR analyses of vanHAXAb and vanYAb in A. balhimycina WT, A. balhimycina ΔvnlRAb, and in A. balhimycina WT overexpressing vnlRAb (WT [vnlRAb]). RNA was isolated at different time points (15/39/63 hr) from the three strains cultivated in balhimycin production medium R5. sigB: transcription of the housekeeping gene sigB. vanHAXAb and vanYAb: transcription of vanHAXAb and vanYAb. For PCR, genomic DNA (gDNA) was used as positive control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5036315&req=5

f3: RT-PCR analyses of vanHAXAb and vanYAb in A. balhimycina WT, A. balhimycina ΔvnlRAb, and in A. balhimycina WT overexpressing vnlRAb (WT [vnlRAb]). RNA was isolated at different time points (15/39/63 hr) from the three strains cultivated in balhimycin production medium R5. sigB: transcription of the housekeeping gene sigB. vanHAXAb and vanYAb: transcription of vanHAXAb and vanYAb. For PCR, genomic DNA (gDNA) was used as positive control.
Mentions: RT-PCR analyses revealed that a vanHAXAb transcript was detectable in A. balhimycina ΔvnlRAb, confirming that the expression of vanHAXAb is independent of vnlRAb (Fig. 3).

View Article: PubMed Central - PubMed

ABSTRACT

In enterococci and in Streptomyces coelicolor, a glycopeptide nonproducer, the glycopeptide resistance genes vanHAX are colocalized with vanRS. The two-component system (TCS) VanRS activates vanHAX transcription upon sensing the presence of glycopeptides. Amycolatopsis balhimycina, the producer of the vancomycin-like glycopeptide balhimycin, also possesses vanHAXAb genes. The genes for the VanRS-like TCS VnlRSAb, together with the carboxypeptidase gene vanYAb, are part of the balhimycin biosynthetic gene cluster, which is located 2 Mb separate from the vanHAXAb. The deletion of vnlRSAb did not affect glycopeptide resistance or balhimycin production. In the A. balhimycina vnlRAb deletion mutant, the vanHAXAb genes were expressed at the same level as in the wild type, and peptidoglycan (PG) analyses proved the synthesis of resistant PG precursors. Whereas vanHAXAb expression in A. balhimycina does not depend on VnlRAb, a VnlRAb-depending regulation of vanYAb was demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR) and RNA-seq analyses. Although VnlRAb does not regulate the vanHAXAb genes in A. balhimycina, its heterologous expression in the glycopeptide-sensitive S. coelicolor ΔvanRSSc deletion mutant restored glycopeptide resistance. VnlRAb activates the vanHAXSc genes even in the absence of VanS. In addition, expression of vnlRAb increases actinorhodin production and influences morphological differentiation in S. coelicolor.

No MeSH data available.