RRC ID 3233
Author Adachi S, Kohiyama M, Onogi T, Hiraga S.
Title Localization of replication forks in wild-type and mukB mutant cells of Escherichia coli.
Journal Mol Genet Genomics
Abstract To examine the subcellular localization of the replication machinery in Escherichia coli, we have developed an immunofluorescence method that allows us to determine the subcellular location of newly synthesized DNA pulse-labeled with 5-bromo-2'-deoxyuridine (BrdU). Using this technique, we have analyzed growing cells. In wild-type cells that showed a single BrdU fluorescence signal, the focus was located in the middle of the cell; in cells with two signals, the foci were localized at positions equivalent to 1/4 and 3/4 of the cell length. The formation of BrdU foci was dependent upon ongoing chromosomal replication. A mutant lacking MukB, which is required for proper partitioning of sister chromosomes, failed to maintain the ordered localization of BrdU foci: (1) a single BrdU focus tended to be localized at a pole-proximal region of the nucleoid, and (2) a focus was often found to consist of two replicating chromosomes. Thus, the positioning of replication forks is affected by the disruption of the mukB gene.
Volume 274(3)
Pages 264-71
Published 2005-10-1
DOI 10.1007/s00438-005-0023-6
PMID 16133165
MeSH Bromodeoxyuridine Chromosomal Proteins, Non-Histone / genetics* DNA / genetics* DNA / metabolism DNA Replication / genetics* Escherichia coli / genetics* Escherichia coli Proteins / genetics* Flow Cytometry Fluorescent Antibody Technique, Indirect / methods Immunohistochemistry
IF 2.879
Times Cited 19
Prokaryotes E. coli YK1100 AZ5372 ME8775(PC2)