TY - JOUR

T1 - The double par locus of virulence factor pB171

T2 - DNA segregation is correlated with oscillation of ParA

AU - Ebersbach, Gitte

AU - Gerdes, Kenn

PY - 2001

Y1 - 2001

N2 - Prokaryotic plasmids and chromosomes encode partitioning (par) loci that segregate DNA to daughter cells before cell division. Recent database analyses showed that almost all known par loci encode an ATPase and a DNA-binding protein, and one or more cis-acting regions where the proteins act. All par-encoded ATPases belong to one of two protein superfamilies, Walker-type and actin-like ATPases. This property was recently used to divide par loci into Types I and II loci. We show here that the Escherichia coli virulence factor pB171 encodes a double par locus that consists of one Type I and one Type II locus. Separately, each locus stabilized a test-plasmid efficiently. Together, the two loci mediated even more efficient plasmid stabilization. The par loci have a unique genetic organization in that they share a common central region at which the two different DNA-binding proteins probably act. Interestingly, a fusion protein consisting of the Walker-type ParA ATPase and Gfp was functional and oscillated in nucleoid regions on a time scale of minutes. ParA-green fluorescent protein (Gfp) oscillation depended on both ParB and parC but was independent of minCDE. Point mutations in the Walker A box motif simultaneously abolished plasmid stabilization and ParA-Gfp oscillation. These observations raise the possibility that ParA oscillation is prerequisite for active plasmid segregation.

AB - Prokaryotic plasmids and chromosomes encode partitioning (par) loci that segregate DNA to daughter cells before cell division. Recent database analyses showed that almost all known par loci encode an ATPase and a DNA-binding protein, and one or more cis-acting regions where the proteins act. All par-encoded ATPases belong to one of two protein superfamilies, Walker-type and actin-like ATPases. This property was recently used to divide par loci into Types I and II loci. We show here that the Escherichia coli virulence factor pB171 encodes a double par locus that consists of one Type I and one Type II locus. Separately, each locus stabilized a test-plasmid efficiently. Together, the two loci mediated even more efficient plasmid stabilization. The par loci have a unique genetic organization in that they share a common central region at which the two different DNA-binding proteins probably act. Interestingly, a fusion protein consisting of the Walker-type ParA ATPase and Gfp was functional and oscillated in nucleoid regions on a time scale of minutes. ParA-green fluorescent protein (Gfp) oscillation depended on both ParB and parC but was independent of minCDE. Point mutations in the Walker A box motif simultaneously abolished plasmid stabilization and ParA-Gfp oscillation. These observations raise the possibility that ParA oscillation is prerequisite for active plasmid segregation.

KW - Bacterial Proteins

KW - Base Sequence

KW - DNA, Bacterial

KW - Green Fluorescent Proteins

KW - Luminescent Proteins

KW - Molecular Sequence Data

KW - Mutation

KW - Plasmids

KW - Recombinant Fusion Proteins

KW - Virulence

U2 - 10.1073/pnas.261569598

DO - 10.1073/pnas.261569598

M3 - Journal article

C2 - 11752455

VL - 98

SP - 15078

EP - 15083

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 - 26

ER -