1887

Chapter 24 : Regulation of Glycopeptide Resistance Genes of Enterococcal Transposon Tn by the VanR-VanS Two-Component Regulatory System

Authors: Michel Arthur1, Florence Depardieu1, Patrice Courvalin1, Theodore Holman2, Zhen Wu2, Gerard Wright2, Christopher T. Walsh2
VIEW AFFILIATIONS HIDE AFFILIATIONS
Affiliations: 1: Unité des Agents Antibacteriéns, Centre National de la Recherche Scientifique, Institut Pasteur, Paris, France; 2: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115
Content Type: Monograph
Publication Year: 1995

Category: Microbial Genetics and Molecular Biology

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Preview this chapter:
Preview Magnify
Zoom in
Zoomout

Regulation of Glycopeptide Resistance Genes of Enterococcal Transposon Tn by the VanR-VanS Two-Component Regulatory System, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555818319/9781555810894_Chap24-1.gif /docserver/preview/fulltext/10.1128/9781555818319/9781555810894_Chap24-2.gif

Abstract:

Glycopeptide antibiotics vancomycin and teicoplanin are used to treat severe infections caused by gram-positive cocci. Strains displaying the so-called VanA resistance phenotype are inducibly resistant to high levels of vancomycin and teicoplanin. Production of the depsipeptide D-Ala-D-Lac is controlled by the VanR-VanS two-component regulatory system that activates transcription of vancomycin resistance genes in response to the presence of glycopeptides in the culture medium. Response regulators (RRs) of this subclass regulate transcription at specific promoters thought to be recognized by the main form of RNA polymerase holoenzyme, corresponding to Eσ in . The first 122 amino acids at the N terminus of VanS are not related in sequence to other HPKs. This region of VanS contains two clusters of hydrophobic amino acids that could correspond to membrane-spanning regions. Validation of the predicted roles of VanS and VanR in sequential phosphoryl group transfer was obtained by overproduction, purification, and assay of the two proteins. The vanR and vanS genes were introduced into the chromosome of a susceptible strain of using an integrative vector. -Activation of transcriptional fusions carried by plasmids were analyzed based on determination of chloramphenicol acetyltransferase (CAT) activity. Mapping of the 5’ end of mRNA by S1 nuclease protection and by primer extension assays identified one transcriptional start site in the vanS-vanH intergenic region. Cloning of , , , , and upstream from the gene in a multicopy vector resulted in high-level transcription of the reporter gene.

Citation: Arthur M, Depardieu F, Courvalin P, Holman T, Wu Z, Wright G, Walsh C. 1995. Regulation of Glycopeptide Resistance Genes of Enterococcal Transposon Tn by the VanR-VanS Two-Component Regulatory System, p 387-391. In Hoch J, Silhavy T (ed), Two-Component Signal Transduction. ASM Press, Washington, DC. doi: 10.1128/9781555818319.ch24

Key Concept Ranking

Glycopeptide Antibiotics
0.7583761
Transcription Initiation
0.5883953
Upstream Promoter
0.5816651
RNA Polymerase
0.5625
0.7583761
Highlighted Text: Show | Hide
Loading full text...

Full text loading...

Figures

Regulation of Glycopeptide Resistance Genes of Enterococcal Transposon Tn1546 by the VanR-VanS Two-Component Regulatory System
[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555818319.chap24-1,webId=/content/author/10.1128/9781555818319.chap24-1,properties={foaf_givenname=Michel, foaf_name=Michel Arthur, foaf_surname=Arthur, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555818319.chap24-aff1]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555818319.chap24-2,webId=/content/author/10.1128/9781555818319.chap24-2,properties={foaf_givenname=Florence, foaf_name=Florence Depardieu, foaf_surname=Depardieu, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555818319.chap24-aff1]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555818319.chap24-3,webId=/content/author/10.1128/9781555818319.chap24-3,properties={foaf_givenname=Patrice, foaf_name=Patrice Courvalin, foaf_surname=Courvalin, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555818319.chap24-aff1]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555818319.chap24-4,webId=/content/author/10.1128/9781555818319.chap24-4,properties={foaf_givenname=Theodore, foaf_name=Theodore Holman, foaf_surname=Holman, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555818319.chap24-aff2]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555818319.chap24-5,webId=/content/author/10.1128/9781555818319.chap24-5,properties={foaf_givenname=Zhen, foaf_name=Zhen Wu, foaf_surname=Wu, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555818319.chap24-aff2]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555818319.chap24-6,webId=/content/author/10.1128/9781555818319.chap24-6,properties={foaf_givenname=Gerard, foaf_name=Gerard Wright, foaf_surname=Wright, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555818319.chap24-aff2]]}], com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/author/10.1128/9781555818319.chap24-7,webId=/content/author/10.1128/9781555818319.chap24-7,properties={foaf_givenname=Christopher T., foaf_name=Christopher T. Walsh, foaf_surname=Walsh, pub_isAffiliatedWith=[com.pub2web.rdf.cci.facet.ContentItem[id=http://asm.metastore.ingenta.com/content/affiliation/10.1128/9781555818319.chap24-aff2]]}]]
Citation: Arthur M, Depardieu F, Courvalin P, Holman T, Wu Z, Wright G, Walsh C. 1995. Regulation of Glycopeptide Resistance Genes of Enterococcal Transposon Tn by the VanR-VanS Two-Component Regulatory System, p 387-391. In Hoch J, Silhavy T (ed), Two-Component Signal Transduction. ASM Press, Washington, DC. doi: 10.1128/9781555818319.ch24
/content/10.1128/9781555818319.chap24.fig24-1
FIGURE 1

Map of Tn and analysis of transcriptional fusions. Open arrows represent coding sequences. Closed and open arrowheads labeled IR and IR. indicate the left and right inverted repeats of the transposon, respectively. Genes , and are cotranscribed from a promoter located in the intergenic region ( ). Restriction sites: B, HI;, II; Bm, I; EI, RI;EV, RV; H, III; He, II;P, I; X, I. The inserts cloned into the multicopy promoter probing vector pAT78 are indicated by solid bars. CAT activity is expressed in nanomoles per minute per milligram of protein. Expression of the cat reporter gene was studied in JH2-2 and in a derivative of JH2-2 harboring a chromosomal copy of . Induction of vancomycin-resistant and -susceptible strains was performed with 50 and 1 µg of vancomycin per milliliter, respectively. R, resistant; S, susceptible; triangles, insertional inactivation of .

10.1128/9781555818319/fig24-1_thmb.gif
10.1128/9781555818319/fig24-1.gif
Image of FIGURE 1
FIGURE 1

Map of Tn and analysis of transcriptional fusions. Open arrows represent coding sequences. Closed and open arrowheads labeled IR and IR. indicate the left and right inverted repeats of the transposon, respectively. Genes , and are cotranscribed from a promoter located in the intergenic region ( ). Restriction sites: B, HI;, II; Bm, I; EI, RI;EV, RV; H, III; He, II;P, I; X, I. The inserts cloned into the multicopy promoter probing vector pAT78 are indicated by solid bars. CAT activity is expressed in nanomoles per minute per milligram of protein. Expression of the cat reporter gene was studied in JH2-2 and in a derivative of JH2-2 harboring a chromosomal copy of . Induction of vancomycin-resistant and -susceptible strains was performed with 50 and 1 µg of vancomycin per milliliter, respectively. R, resistant; S, susceptible; triangles, insertional inactivation of .

Citation: Arthur M, Depardieu F, Courvalin P, Holman T, Wu Z, Wright G, Walsh C. 1995. Regulation of Glycopeptide Resistance Genes of Enterococcal Transposon Tn by the VanR-VanS Two-Component Regulatory System, p 387-391. In Hoch J, Silhavy T (ed), Two-Component Signal Transduction. ASM Press, Washington, DC. doi: 10.1128/9781555818319.ch24
Permissions and Reprints Request Permissions
Download as Powerpoint

References

/content/book/10.1128/9781555818319.chap24
1. Al-Obeid, S.,, L. Gutmann,, D. M. Shlaes,, R. Williamson,, and E. Collatz. 1990. Comparison of vancomycin-inducible proteins from four strains of Enterococci. FEMS Microbiol. Lett. 70: 101– 106.
2. Arthur, M.,, and P. Courvalin. 1993. Genetics and mechanisms of glycopeptide resistance in enterococci. Antimicrob. Agents Chemother. 37: 1563– 1571.
3. Arthur, M.,, C. Molinas,, and P. Courvalin. 1992a. Sequence of the vanY gene required for production of a vancomycin-inducible D.D-carboxypeptidase in Enterococcus faecium BM4147. Gene 120: 111– 114
4. Arthur, M.,, C. Molinas,, and P. Courvalin. 1992b. The VanS-VanR two-component regulatory system controls synthesis of depsipeptide peptidoglycan precursors in Enterococcus faecium BM4147. J. Bacteriol 174: 2582– 2591.
5. Arthur, M.,, C. Molinas,, F. Depardieu,, and P. Courvalin. 1993. Characterization of Tnl546, a Tn3-related transposon conferring glycopeptide resistance by synthesis of depsipeptide peptidoglycan precursors in Enterococcus faecium BM4147. J. Bacteriol. 175: 117– 127.
6. Bugg, T. D. H.,, G. D. Wright,, S. Dutka-Malen,, M. Arthur,, P. Courvalin,, and C. T. Walsh. 1991. Molecular basis for vancomycin resistance in Enterococcus faecium BM4147: biosynthesis of a depsipeptide peptidoglycan precursor by vancomycin resistance proteins VanH and VanA. Biochemistry 30: 10408– 10415.
7. Dutka-Malen, S.,, C. Molinas,, M. Arthur,, and P. Courvalin. 1990. The VANA glycopeptide resistance protein is related to D-alanyl-D-alanine ligase cell wall biosynthesis enzymes. Mol. Gen. Genet. 224: 364– 372.
8. Gutmann, L.,, D. Billot-Klein,, S. Al-Obeid,, I. Klare,, S. Francoual,, E. Collatz,, and J. Van Heijenoort. 1992. Inducible carboxypeptidase activity in vancomycin-resistant enterococci. Antimicrob. Agents Chemother. 36: 77– 80.
9. Handwerger, S.,, and A. Kolokathis. 1990. Induction of vancomycin resistance in Enterococcus faecium by inhibition of transglycosylation. FEMS Microbiol. Lett. 70: 167– 170.
10. Hess, J. E.,, R. B. Bourret,, and M. I. Simon. 1988. Histidine phosphorylation and phosphoryl group transfer in bacterial chemotaxis. Nature (London) 336: 139– 143.
11. Holman, T. R.,, Z. Wu,, B. L. Wanner,, and C. T. Walsh. 1994. Identification of the DNA-binding site for the phosphorylated VanR protein required for vancomycin resistance in Entococcus faecium. Biochemistry 33: 4625– 4631.
12. Igo, M. M.,, A. J. Ninfa,, J. B. Stock,, and T. J. Silhavy. 1989. Phosphorylation and dephosphorylation of a bacterial transcriptional activator by a transmembrane receptor. Genes Dev. 3: 1725– 1734.
13. Johnson, A. P.,, A. H. C. Uttley,, N. Woodford,, and R. C. George. 1990. Resistance to vancomycin and teicoplanin: an emerging clinical problem. Clin. Microbiol. Rev. 3: 280– 291.
14. Leclercq, R.,, E. Derlot,, M. Weber,, J. Duval,, and P. Courvalin. 1989. Transferable vancomycin and teicoplanin resistance in Enterococcus faecium. Antimicrob. Agents Chemother. 33: 10– 15.
15. Murray, B. E. 1990. The life and times of the Enterococcus. Clin. Microbiol. Rev. 3: 46– 65.
16. Reynolds, P. E. 1989. Structure, biochemistry and mechanism of action of glycopeptide antibiotics. Eur. J. Clin. Microbiol. Infect. Dis. 8: 943– 950.
17. Shlaes, D. M.,, A. Bouvet,, C. Devine,, J. H. Shlaes,, S. Al-Obeid,, and R. Williamson. 1989. Inducible, transferable resistance to vancomycin in Enterococcus faecalis A256. Antimicrob. Agents Chemother. 33: 198– 203.
18. Stock, J. B.,, A. J. Ninfa,, and A. M. Stock. 1989. Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol. Rev. 53: 450– 490.
19. Williamson, R. C.,, S. Al-Obeid,, J. H. Shlaes,, F. W. Goldstein,, and D. M. Shlaes. 1989. Inducible resistance to vancomycin in Enterococcus faecium D366J. Infect. Dis. 159: 1095– 1104.
20. Wright, G. D.,, T. R. Holman,, and C. T. Walsh. 1993. Purification and characterization of VanR and the cytosolic domain of VanS: a two-component regulatory system required for vancomycin resistance in Enterococcus faecium BM4147. Biochemistry 32: 5057– 5063.

Back to top
This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error