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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.


(A, B) EMBOSS stretcher pairwise sequence alignment of VnlRSAb and VanRSSc. (A) EMBOSS stretcher pairwise sequence alignment of VnlSAb and VanSSc. Transmembrane domains are indicated in blue. The extracytosolic domain is highlighted by a red box. (B) EMBOSS stretcher pairwise sequence alignment of VnlRAb and VanRSc. The site of aspartate phosphorylation is indicated by red and that of the proposed autophosphorylation by green arrow. (C) Organization of the resistance genes in Streptomyces coelicolor compared with that of Amycolatopsis balhimycina. “-” for a mismatch or a gap; “.” for any small positive score; “:” for a similarity, which scores more than 1.0; and “I” for an identity where both sequences have the same residue.
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f1: (A, B) EMBOSS stretcher pairwise sequence alignment of VnlRSAb and VanRSSc. (A) EMBOSS stretcher pairwise sequence alignment of VnlSAb and VanSSc. Transmembrane domains are indicated in blue. The extracytosolic domain is highlighted by a red box. (B) EMBOSS stretcher pairwise sequence alignment of VnlRAb and VanRSc. The site of aspartate phosphorylation is indicated by red and that of the proposed autophosphorylation by green arrow. (C) Organization of the resistance genes in Streptomyces coelicolor compared with that of Amycolatopsis balhimycina. “-” for a mismatch or a gap; “.” for any small positive score; “:” for a similarity, which scores more than 1.0; and “I” for an identity where both sequences have the same residue.

Mentions: Streptomyces coelicolor A3(2) is neither a pathogen nor a glycopeptide producer, but it is likely to encounter glycopeptides in its natural habitat. Therefore, it benefits from carrying vanRSSc, vanHAXSc, vanKSc, and vanJSc (Fig. 1C). VanHSc is a d-stereospecific lactate dehydrogenase that converts pyruvate to d-Lac. VanASc is a d-Ala-d-Ala-ligase family protein that ligates d-Ala and d-Lac to d-Ala-d-Lac-depsipeptides. VanXSc is a highly selective carboxypeptidase that cleaves the remaining d-Ala-d-Ala-dipeptide. VanKSc belongs to the Fem family of enzymes, which add the cross-bridging amino acid(s) to the stem pentapeptide of PG precursors.19 VanY is a membrane protein conferring resistance to teicoplanin. To identify the precise nature of the ligand signal that activates glycopeptide resistance in S. coelicolor A3(2), the VanB-type HK VanSSc, sensing vancomycin, but not teicoplanin, was investigated.21,22 Investigation on VanSSc revealed opposed results. On the one hand, it was shown by cross-linking experiments that vancomycin is the direct ligand of the VanSSc.21 On the other hand, Kwun et al.22 demonstrated that VanSSc is activated by vancomycin in complex with the d-Ala-d-Ala termini of PG precursors.


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
(A, B) EMBOSS stretcher pairwise sequence alignment of VnlRSAb and VanRSSc. (A) EMBOSS stretcher pairwise sequence alignment of VnlSAb and VanSSc. Transmembrane domains are indicated in blue. The extracytosolic domain is highlighted by a red box. (B) EMBOSS stretcher pairwise sequence alignment of VnlRAb and VanRSc. The site of aspartate phosphorylation is indicated by red and that of the proposed autophosphorylation by green arrow. (C) Organization of the resistance genes in Streptomyces coelicolor compared with that of Amycolatopsis balhimycina. “-” for a mismatch or a gap; “.” for any small positive score; “:” for a similarity, which scores more than 1.0; and “I” for an identity where both sequences have the same residue.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: (A, B) EMBOSS stretcher pairwise sequence alignment of VnlRSAb and VanRSSc. (A) EMBOSS stretcher pairwise sequence alignment of VnlSAb and VanSSc. Transmembrane domains are indicated in blue. The extracytosolic domain is highlighted by a red box. (B) EMBOSS stretcher pairwise sequence alignment of VnlRAb and VanRSc. The site of aspartate phosphorylation is indicated by red and that of the proposed autophosphorylation by green arrow. (C) Organization of the resistance genes in Streptomyces coelicolor compared with that of Amycolatopsis balhimycina. “-” for a mismatch or a gap; “.” for any small positive score; “:” for a similarity, which scores more than 1.0; and “I” for an identity where both sequences have the same residue.
Mentions: Streptomyces coelicolor A3(2) is neither a pathogen nor a glycopeptide producer, but it is likely to encounter glycopeptides in its natural habitat. Therefore, it benefits from carrying vanRSSc, vanHAXSc, vanKSc, and vanJSc (Fig. 1C). VanHSc is a d-stereospecific lactate dehydrogenase that converts pyruvate to d-Lac. VanASc is a d-Ala-d-Ala-ligase family protein that ligates d-Ala and d-Lac to d-Ala-d-Lac-depsipeptides. VanXSc is a highly selective carboxypeptidase that cleaves the remaining d-Ala-d-Ala-dipeptide. VanKSc belongs to the Fem family of enzymes, which add the cross-bridging amino acid(s) to the stem pentapeptide of PG precursors.19 VanY is a membrane protein conferring resistance to teicoplanin. To identify the precise nature of the ligand signal that activates glycopeptide resistance in S. coelicolor A3(2), the VanB-type HK VanSSc, sensing vancomycin, but not teicoplanin, was investigated.21,22 Investigation on VanSSc revealed opposed results. On the one hand, it was shown by cross-linking experiments that vancomycin is the direct ligand of the VanSSc.21 On the other hand, Kwun et al.22 demonstrated that VanSSc is activated by vancomycin in complex with the d-Ala-d-Ala termini of PG precursors.

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.