| 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 |
2.797
|
| Times Cited |
19
|
|
WOS Category
|
GENETICS & HEREDITY
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Prokaryotes E. coli |
YK1100
AZ5372
ME8775(PC2) |
