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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.


Growth and resistance of the S. coelicolor M600 ΔvanRSSc complemented with different combinations of vnlRSAb. (A) Growth on YM agar containing no antibiotic. (B) Growth on YM agar containing apramycin (100 mg/ml) (Apra 100) to prove plasmid integration. (C) Growth on YM agar containing balhimycin (10 mg/ml) (Bal 10). (D) Growth on YM agar containing teicoplanin (10 mg/ml) (Teico 10). (E) Growth on YM agar containing no antibiotic. M600, S. coelicolor M600.
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f4: Growth and resistance of the S. coelicolor M600 ΔvanRSSc complemented with different combinations of vnlRSAb. (A) Growth on YM agar containing no antibiotic. (B) Growth on YM agar containing apramycin (100 mg/ml) (Apra 100) to prove plasmid integration. (C) Growth on YM agar containing balhimycin (10 mg/ml) (Bal 10). (D) Growth on YM agar containing teicoplanin (10 mg/ml) (Teico 10). (E) Growth on YM agar containing no antibiotic. M600, S. coelicolor M600.

Mentions: Introduction of vnlRSAb and of vnlRAb alone into S. coelicolor M600 ΔvanRSSc resulted in balhimycin-resistant strains (Fig. 4). In contrast, expression of vnlSAb alone did not change the glycopeptide-sensitive phenotype of the S. coelicolor ΔvanRSSc mutant. These results indicated that VnlRAb from A. balhimycina is able to activate the transcription of vanHAXSc in S. coelicolor M600 also in the absence of VnlSAb. Since in S. coelicolor M600 VanRSc∼P can be generated in a VanSSc-independent manner using acetylphosphate,6 we suggest a similar activation of VnlRAb in the absence of VnlSAb or VanSSc.


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
Growth and resistance of the S. coelicolor M600 ΔvanRSSc complemented with different combinations of vnlRSAb. (A) Growth on YM agar containing no antibiotic. (B) Growth on YM agar containing apramycin (100 mg/ml) (Apra 100) to prove plasmid integration. (C) Growth on YM agar containing balhimycin (10 mg/ml) (Bal 10). (D) Growth on YM agar containing teicoplanin (10 mg/ml) (Teico 10). (E) Growth on YM agar containing no antibiotic. M600, S. coelicolor M600.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Growth and resistance of the S. coelicolor M600 ΔvanRSSc complemented with different combinations of vnlRSAb. (A) Growth on YM agar containing no antibiotic. (B) Growth on YM agar containing apramycin (100 mg/ml) (Apra 100) to prove plasmid integration. (C) Growth on YM agar containing balhimycin (10 mg/ml) (Bal 10). (D) Growth on YM agar containing teicoplanin (10 mg/ml) (Teico 10). (E) Growth on YM agar containing no antibiotic. M600, S. coelicolor M600.
Mentions: Introduction of vnlRSAb and of vnlRAb alone into S. coelicolor M600 ΔvanRSSc resulted in balhimycin-resistant strains (Fig. 4). In contrast, expression of vnlSAb alone did not change the glycopeptide-sensitive phenotype of the S. coelicolor ΔvanRSSc mutant. These results indicated that VnlRAb from A. balhimycina is able to activate the transcription of vanHAXSc in S. coelicolor M600 also in the absence of VnlSAb. Since in S. coelicolor M600 VanRSc∼P can be generated in a VanSSc-independent manner using acetylphosphate,6 we suggest a similar activation of VnlRAb in the absence of VnlSAb or VanSSc.

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.