PMID:16133165
| Citation |
Adachi, S, Kohiyama, M, Onogi, T and Hiraga, S (2005) Localization of replication forks in wild-type and mukB mutant cells of Escherichia coli. Mol. Genet. Genomics 274:264-71 |
|---|---|
| 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. |
| Links |
PubMed Online version:10.1007/s00438-005-0023-6 |
| Keywords |
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 |
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