RRC ID |
30153
|
著者 |
Onogi T, Niki H, Yamazoe M, Hiraga S.
|
タイトル |
The assembly and migration of SeqA-Gfp fusion in living cells of Escherichia coli.
|
ジャーナル |
Mol Microbiol
|
Abstract |
SeqA protein, which binds to hemi-methylated GATC sequences of DNA, is localized to discrete fluorescent foci in wild-type Escherichia coli cells. In this work, we observed cellular localization of the SeqA-Gfp fusion in living cells. SeqA-Gfp was localized to a discrete focus or foci in wild-type and seqA null mutant cells, but the fusion was dispersed in the whole cell in dam null mutant cells lacking Dam methyltransferase. These results were consistent with the previous description of the localization of SeqA by immunofluorescence microscopy. Time-lapse experiments revealed that duplicated SeqA-Gfp foci migrated rapidly in opposite directions. Flow cytometry demonstrated that the fusion restored synchronous replication of chromosomal DNA from multiple origins in seqA null mutant cells, indicating that SeqA-Gfp is biologically active. Immunoprecipitation of the fusion from cell extracts using anti-Gfp antibody indicated that the fusion was assembled with the wild-type SeqA protein.
|
巻・号 |
31(6)
|
ページ |
1775-82
|
公開日 |
1999-3-1
|
DOI |
10.1046/j.1365-2958.1999.01313.x
|
PMID |
10209749
|
MeSH |
Bacterial Outer Membrane Proteins
Bacterial Proteins / metabolism*
Cell Nucleus / physiology
Chromosomal Proteins, Non-Histone*
DNA Replication / physiology
DNA-Binding Proteins
Electrophoresis, Polyacrylamide Gel
Escherichia coli / cytology
Escherichia coli / physiology*
Escherichia coli Proteins*
Flow Cytometry
Green Fluorescent Proteins
Luminescent Proteins / metabolism*
Models, Genetic
Mutagenesis
Plasmids
Precipitin Tests
Recombinant Fusion Proteins / metabolism*
Time Factors
Transcription Factors*
|
IF |
3.418
|
引用数 |
91
|
WOS 分野
|
MICROBIOLOGY
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
原核生物(大腸菌) |
ME7825
ME7826
ME7827 |