Centromere anatomy in the multidrug-resistant pathogen Enterococcus faecium

Andrew Derome; Christian Hoischen; Malte Bussiek; Ruth Grady; Malgorzata Adamczyk; Barbara Kedzierska; Stephan Diekmann; Daniela Barilla; Finbarr Hayes

doi:10.1073/pnas.0704681105

Centromere anatomy in the multidrug-resistant pathogen Enterococcus faecium

Andrew Derome, Christian Hoischen, Malte Bussiek, Ruth Grady, Malgorzata Adamczyk, Barbara Kedzierska, Stephan Diekmann, Daniela Barilla, Finbarr Hayes

Biology

Research output: Contribution to journal › Article › peer-review

Abstract

Multidrug-resistant variants of the opportunistic human pathogen Enterococcus have recently emerged as leading agents of nosocomial infection. The acquisition of plasmid-borne resistance genes is a driving force in antibiotic-resistance evolution in enterococci. The segregation locus of a high-level gentamicin-resistance plasmid, pGENT, in Enterococcus faecium, was identified and dissected. This locus includes overlapping genes encoding PrgP, a member of the ParA superfamily of segregation proteins, and PrgO, a site-specific DNA binding homodimer that recognizes the cenE centromere upstream of prgPO. The centromere has a distinctive organization comprising three subsites, CESII separates CESI and CESIII, each of which harbors seven TATA boxes spaced by half-helical turns. PrgO independently binds both CESI and CESIII, but with different affinities. The topography of the complex was probed by atomic force microscopy, revealing discrete PrgO foci positioned asymmetrically at the CESI and CESIII subsites. Bending analysis demonstrated that cenE is intrinsically curved. The organization of the cenE site and of certain other plasmid centromeres mirrors that of yeast centromeres, which may reflect a common architectural requirement during assembly of the mitotic apparatus in yeast and bacteria. Moreover, segregation modules homologous to that of pGENT are widely disseminated on vancomycin and other resistance plasmids in enterococci. An improved understanding of segrosome assembly may highlight new interventions geared toward combating antibiotic resistance in these insidious pathogens.

Original language	English
Pages (from-to)	2151-2156
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	105
Issue number	6
DOIs	https://doi.org/10.1073/pnas.0704681105
Publication status	Published - 12 Feb 2008

Keywords

antibiotic resistance
plasmid
segregation
partition
gentamicin
BACTERIAL-DNA SEGREGATION
CURVED DNA
PLASMID CENTROMERE
SALT DEPENDENCE
GEL MIGRATION
PROTEIN
CURVATURE
PARF
IDENTIFICATION
TEMPERATURE

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10.1073/pnas.0704681105

http://www.pnas.org/content/105/6/2151

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title = "Centromere anatomy in the multidrug-resistant pathogen Enterococcus faecium",

abstract = "Multidrug-resistant variants of the opportunistic human pathogen Enterococcus have recently emerged as leading agents of nosocomial infection. The acquisition of plasmid-borne resistance genes is a driving force in antibiotic-resistance evolution in enterococci. The segregation locus of a high-level gentamicin-resistance plasmid, pGENT, in Enterococcus faecium, was identified and dissected. This locus includes overlapping genes encoding PrgP, a member of the ParA superfamily of segregation proteins, and PrgO, a site-specific DNA binding homodimer that recognizes the cenE centromere upstream of prgPO. The centromere has a distinctive organization comprising three subsites, CESII separates CESI and CESIII, each of which harbors seven TATA boxes spaced by half-helical turns. PrgO independently binds both CESI and CESIII, but with different affinities. The topography of the complex was probed by atomic force microscopy, revealing discrete PrgO foci positioned asymmetrically at the CESI and CESIII subsites. Bending analysis demonstrated that cenE is intrinsically curved. The organization of the cenE site and of certain other plasmid centromeres mirrors that of yeast centromeres, which may reflect a common architectural requirement during assembly of the mitotic apparatus in yeast and bacteria. Moreover, segregation modules homologous to that of pGENT are widely disseminated on vancomycin and other resistance plasmids in enterococci. An improved understanding of segrosome assembly may highlight new interventions geared toward combating antibiotic resistance in these insidious pathogens.",

keywords = "antibiotic resistance, plasmid, segregation, partition, gentamicin, BACTERIAL-DNA SEGREGATION, CURVED DNA, PLASMID CENTROMERE, SALT DEPENDENCE, GEL MIGRATION, PROTEIN, CURVATURE, PARF, IDENTIFICATION, TEMPERATURE",

author = "Andrew Derome and Christian Hoischen and Malte Bussiek and Ruth Grady and Malgorzata Adamczyk and Barbara Kedzierska and Stephan Diekmann and Daniela Barilla and Finbarr Hayes",

year = "2008",

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doi = "10.1073/pnas.0704681105",

language = "English",

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T1 - Centromere anatomy in the multidrug-resistant pathogen Enterococcus faecium

AU - Derome, Andrew

AU - Hoischen, Christian

AU - Bussiek, Malte

AU - Grady, Ruth

AU - Adamczyk, Malgorzata

AU - Kedzierska, Barbara

AU - Diekmann, Stephan

AU - Barilla, Daniela

AU - Hayes, Finbarr

PY - 2008/2/12

Y1 - 2008/2/12

N2 - Multidrug-resistant variants of the opportunistic human pathogen Enterococcus have recently emerged as leading agents of nosocomial infection. The acquisition of plasmid-borne resistance genes is a driving force in antibiotic-resistance evolution in enterococci. The segregation locus of a high-level gentamicin-resistance plasmid, pGENT, in Enterococcus faecium, was identified and dissected. This locus includes overlapping genes encoding PrgP, a member of the ParA superfamily of segregation proteins, and PrgO, a site-specific DNA binding homodimer that recognizes the cenE centromere upstream of prgPO. The centromere has a distinctive organization comprising three subsites, CESII separates CESI and CESIII, each of which harbors seven TATA boxes spaced by half-helical turns. PrgO independently binds both CESI and CESIII, but with different affinities. The topography of the complex was probed by atomic force microscopy, revealing discrete PrgO foci positioned asymmetrically at the CESI and CESIII subsites. Bending analysis demonstrated that cenE is intrinsically curved. The organization of the cenE site and of certain other plasmid centromeres mirrors that of yeast centromeres, which may reflect a common architectural requirement during assembly of the mitotic apparatus in yeast and bacteria. Moreover, segregation modules homologous to that of pGENT are widely disseminated on vancomycin and other resistance plasmids in enterococci. An improved understanding of segrosome assembly may highlight new interventions geared toward combating antibiotic resistance in these insidious pathogens.

AB - Multidrug-resistant variants of the opportunistic human pathogen Enterococcus have recently emerged as leading agents of nosocomial infection. The acquisition of plasmid-borne resistance genes is a driving force in antibiotic-resistance evolution in enterococci. The segregation locus of a high-level gentamicin-resistance plasmid, pGENT, in Enterococcus faecium, was identified and dissected. This locus includes overlapping genes encoding PrgP, a member of the ParA superfamily of segregation proteins, and PrgO, a site-specific DNA binding homodimer that recognizes the cenE centromere upstream of prgPO. The centromere has a distinctive organization comprising three subsites, CESII separates CESI and CESIII, each of which harbors seven TATA boxes spaced by half-helical turns. PrgO independently binds both CESI and CESIII, but with different affinities. The topography of the complex was probed by atomic force microscopy, revealing discrete PrgO foci positioned asymmetrically at the CESI and CESIII subsites. Bending analysis demonstrated that cenE is intrinsically curved. The organization of the cenE site and of certain other plasmid centromeres mirrors that of yeast centromeres, which may reflect a common architectural requirement during assembly of the mitotic apparatus in yeast and bacteria. Moreover, segregation modules homologous to that of pGENT are widely disseminated on vancomycin and other resistance plasmids in enterococci. An improved understanding of segrosome assembly may highlight new interventions geared toward combating antibiotic resistance in these insidious pathogens.

KW - antibiotic resistance

KW - plasmid

KW - segregation

KW - partition

KW - gentamicin

KW - BACTERIAL-DNA SEGREGATION

KW - CURVED DNA

KW - PLASMID CENTROMERE

KW - SALT DEPENDENCE

KW - GEL MIGRATION

KW - PROTEIN

KW - CURVATURE

KW - PARF

KW - IDENTIFICATION

KW - TEMPERATURE

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U2 - 10.1073/pnas.0704681105

DO - 10.1073/pnas.0704681105

M3 - Article

VL - 105

SP - 2151

EP - 2156

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 6

ER -

Centromere anatomy in the multidrug-resistant pathogen Enterococcus faecium

Abstract

Keywords

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