PMID:9473485

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Citation

Kurokawa, K, Mizushima, T, Kubota, T, Tsuchiya, T, Katayama, T and Sekimizu, K (1998) A stimulation factor for hydrolysis of ATP bound to DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli. Biochem. Biophys. Res. Commun. 243:90-5

Abstract

Hydrolysis of ATP bound to DnaA protein by its intrinsic ATPase activity negatively controls chromosomal DNA replication in Escherichia coli. We developed a new in vitro assay system for ATP hydrolysis, which makes feasible a search for factors affecting the ATPase activity of DnaA protein. A crude cell extract enhanced the hydrolysis of ATP bound to DnaA protein, in a dose-dependent manner. Gel-filtration analyses revealed a single entity of the stimulation factor for the ATP hydrolysis and an apparent molecular mass of 170 kDa. The stimulation activity for ATP hydrolysis coeluted with the inactivation activity for DnaA protein initiating an oriC DNA replication, as determined by anion-exchange and gel-filtration column chromatographies. Activity of the stimulation factor required DNA and ATP. These observations suggested that IdaA protein, a previously described negative factor for DnaA protein, inactivated DnaA protein through stimulation of the hydrolysis of ATP bound to DnaA protein.

Links

PubMed Online version:10.1006/bbrc.1997.8069

Keywords

Adenosine Diphosphate/metabolism; Adenosine Triphosphate/isolation & purification; Adenosine Triphosphate/metabolism; Bacterial Proteins/chemistry; Bacterial Proteins/isolation & purification; Bacterial Proteins/metabolism; DNA Replication; DNA, Bacterial/biosynthesis; DNA-Binding Proteins/chemistry; DNA-Binding Proteins/isolation & purification; DNA-Binding Proteins/metabolism; Escherichia coli/genetics; Escherichia coli/metabolism; Hydrolysis; Molecular Weight; Precipitin Tests; Protein Binding; Replication Origin

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