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A Novel Cytoplasmic Hemimethylated oriC Binding Activity
- Source :
- Journal of Biological Chemistry. 271:7404-7411
- Publication Year :
- 1996
- Publisher :
- Elsevier BV, 1996.
-
Abstract
- Using hemimethylated, fully methylated, and unmethylated oligonucleotide probes corresponding to part of the origin of Escherichia coli DNA replication, oriC (+81-136), we have characterized a novel hemimethylated DNA-specific protein binding activity. This activity appears to be located in the cytoplasm rather than in membrane fractions. It has been partially purified and, in DNase footprinting analysis, found to preferentially protect only a subset of the hemimethylated GATC sites present in the minimal oriC. These sites are found adjacent to the DnaA binding box, R1, and overlap the integration host factor binding site. The activity does not correspond to known hemimethylated binding proteins, although in the seqA deletion mutant, there is a 3-fold reduction of the activity. The stage of the cell cycle in synchronized PC2 cultures does not seem to significantly affect the relative levels of this binding activity. A possible role in sequestration of the newly replicated hemimethylated origin is discussed.
- Subjects :
- DNA Replication
DNA, Bacterial
Cytoplasm
Molecular Sequence Data
DNA Footprinting
DNA footprinting
Replication Origin
Plasma protein binding
Biology
Binding, Competitive
Methylation
Biochemistry
DNA-binding protein
Bacterial Proteins
SeqA protein domain
Escherichia coli
Binding site
Molecular Biology
Sequence Deletion
Deoxyribonucleases
Base Sequence
Oligonucleotide
Escherichia coli Proteins
Cell Biology
Chromatography, Ion Exchange
DnaA
DNA-Binding Proteins
Kinetics
Mutagenesis
Chromatography, Gel
Electrophoresis, Polyacrylamide Gel
Oligonucleotide Probes
Bacterial Outer Membrane Proteins
Transcription Factors
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 271
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....5f8b566a035802c348cc188814da9a66