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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.

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RT-PCR analyses of S. coelicolor M600 and different S. coelicolor M600 mutants. RNA was isolated after 25 hr of cultivation in the absence of any glycopeptide. hrdB: transcription of the housekeeping gene hrdBSc. vanSSc, vanRSc, and vanHAXSc: transcription of vanSSc, vanRSc, and vanHAXSc. For PCR, genomic DNA (gDNA) was used as positive control.
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f5: RT-PCR analyses of S. coelicolor M600 and different S. coelicolor M600 mutants. RNA was isolated after 25 hr of cultivation in the absence of any glycopeptide. hrdB: transcription of the housekeeping gene hrdBSc. vanSSc, vanRSc, and vanHAXSc: transcription of vanSSc, vanRSc, and vanHAXSc. For PCR, genomic DNA (gDNA) was used as positive control.

Mentions: The activation of the vanHAXSc genes in the complemented S. coelicolor M600 ΔvanRSSc mutant with vnlRSAb and vnlRAb was further analyzed by RT-PCR. For this purpose, RNA was isolated from 25-hr-old liquid cultures grown without addition of any glycopeptide. A vanHAXSc transcript was detected when S. coelicolor M600 ΔvanRSSc was complemented with vnlRSAb or with vnlRAb alone. However, in S. coelicolor M600 and in the S. coelicolor M600 ΔvanRSSc mutant, transcription of the vanHAXSc failed (Fig. 5), confirming the functionality of VnlRAb as transcriptional activator in S. coelicolor.


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 S. coelicolor M600 and different S. coelicolor M600 mutants. RNA was isolated after 25 hr of cultivation in the absence of any glycopeptide. hrdB: transcription of the housekeeping gene hrdBSc. vanSSc, vanRSc, and vanHAXSc: transcription of vanSSc, vanRSc, and vanHAXSc. 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

f5: RT-PCR analyses of S. coelicolor M600 and different S. coelicolor M600 mutants. RNA was isolated after 25 hr of cultivation in the absence of any glycopeptide. hrdB: transcription of the housekeeping gene hrdBSc. vanSSc, vanRSc, and vanHAXSc: transcription of vanSSc, vanRSc, and vanHAXSc. For PCR, genomic DNA (gDNA) was used as positive control.
Mentions: The activation of the vanHAXSc genes in the complemented S. coelicolor M600 ΔvanRSSc mutant with vnlRSAb and vnlRAb was further analyzed by RT-PCR. For this purpose, RNA was isolated from 25-hr-old liquid cultures grown without addition of any glycopeptide. A vanHAXSc transcript was detected when S. coelicolor M600 ΔvanRSSc was complemented with vnlRSAb or with vnlRAb alone. However, in S. coelicolor M600 and in the S. coelicolor M600 ΔvanRSSc mutant, transcription of the vanHAXSc failed (Fig. 5), confirming the functionality of VnlRAb as transcriptional activator in S. coelicolor.

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.


Related in: MedlinePlus