Replication cycle-coordinated change of the adenine nucleotide-bound forms of DnaA protein in Escherichia coli

• Published in: The EMBO journal 1999-12, Vol.18 (23), p.6642-6652
• Main Authors: Kurokawa, Kenji, Nishida, Satoshi, Emoto, Akiko, Sekimizu, Kazuhisa, Katayama, Tsutomu
• Format: Article
• Language: English
• Subjects: Adenine - chemistry; Adenosine diphosphate; Adenosine Diphosphate - metabolism; Adenosine Triphosphate - metabolism; ADP; ATP; Bacterial Outer Membrane Proteins; Bacterial Proteins - biosynthesis; Bacterial Proteins - chemistry; Bacterial Proteins - metabolism; Cell Cycle; Dam protein; Deoxyribonucleic acid; DNA; DNA polymerase; DNA Replication; DNA-Binding Proteins - biosynthesis; DNA-Binding Proteins - chemistry; DNA-Directed DNA Polymerase; DnaA; DnaA protein; Dose-Response Relationship, Drug; E coli; Escherichia coli; Escherichia coli - chemistry; Escherichia coli - metabolism; Escherichia coli Proteins; Hydrolysis; Precipitin Tests; Protein Binding; Protein synthesis; Replication Origin; SeqA protein; sliding clamp; Temperature; Time Factors; Transcription Factors
• ISSN: 0261-4189; 1460-2075; 1460-2075
• DOI: 10.1093/emboj/18.23.6642

The ATP‐bound but not the ADP‐bound form of DnaA protein is active for replication initiation at the Escherichia coli chromosomal origin. The hydrolysis of ATP bound to DnaA is accelerated by the sliding clamp of DNA polymerase III loaded on DNA. Using a culture of randomly dividing cells, we now have evidence that the cellular level of ATP–DnaA is repressed to only ∼20% of the total DnaA molecules, in a manner depending on DNA replication. In a synchronized culture, the ATP–DnaA level showed oscillation that has a temporal increase around the time of initiation, and decreases rapidly after initiation. Production of ATP–DnaA depended on concomitant protein synthesis, but not on SOS response, Dam or SeqA. Regeneration of ATP–DnaA from ADP–DnaA was also observed. These results indicate that the nucleotide form shifts of DnaA are tightly linked with an epistatic cell cycle event and with the chromosomal replication system.