RRC ID 3233
著者 Adachi S, Kohiyama M, Onogi T, Hiraga S.
タイトル Localization of replication forks in wild-type and mukB mutant cells of Escherichia coli.
ジャーナル 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.
巻・号 274(3)
ページ 264-71
公開日 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 2.797
引用数 19
WOS 分野 GENETICS & HEREDITY BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
原核生物(大腸菌) YK1100 AZ5372 ME8775(PC2)