A novel mode of DnaA-DnaA interaction promotes ADP dissociation for reactivation of replication initiation activity

Ryo Sugiyama, Kazutoshi Kasho, Kenya Miyoshi, Shogo Ozaki, Wataru Kagawa, Hitoshi Kurumizaka, Tsutomu Katayama

Research output: Contribution to journalArticlepeer-review

Abstract

ATP-DnaA is temporally increased to initiate replication during the cell cycle. Two chromosomal loci, DARS (DnaA-reactivating sequences) 1 and 2, promote ATP-DnaA production by nucleotide exchange of ADP-DnaA for timely initiation. ADP-DnaA complexes are constructed on DARS1 and DARS2, bearing a cluster of three DnaA-binding sequences (DnaA boxes I-III), promoting ADP dissociation. Although DnaA has an AAA+ domain, which ordinarily directs construction of oligomers in a head-to-tail manner, DnaA boxes I and II are oriented oppositely. In this study, we constructed a structural model of a head-to-head dimer of DnaA AAA+ domains, and analyzed residues residing on the interface of the model dimer. Gln208 was specifically required for DARS-dependent ADP dissociation in vitro, and in vivo analysis yielded consistent results. Additionally, ADP release from DnaA protomers bound to DnaA boxes I and II was dependent on Gln208 of the DnaA protomers, and DnaA box III-bound DnaA did not release ADP nor require Gln208 for ADP dissociation by DARS-DnaA complexes. Based on these and other findings, we propose a model for DARS-DnaA complex dynamics during ADP dissociation, and provide novel insight into the regulatory mechanisms of DnaA and the interaction modes of AAA+ domains.

Original languageEnglish
Pages (from-to)11209-11224
Number of pages16
JournalNucleic acids research
Volume47
Issue number21
DOIs
Publication statusPublished - Dec 2 2019

All Science Journal Classification (ASJC) codes

  • Genetics

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