||Kazutoshi Kasho, Lukas Krasauskas, Vytautas Smirnovas, Gorazd Stojkovič, Ludmilla A Morozova-Roche, Sjoerd Wanrooij, Human Polymerase δ-Interacting Protein 2 (PolDIP2) Inhibits the Formation of Human Tau Oligomers and Fibrils., International journal of molecular sciences, 10.3390/ijms22115768, 22, 11, 2021.05.
||Kazutoshi Kasho, Gorazd Stojkovič, Cristina Velázquez-Ruiz, Maria Isabel Martínez-Jiménez, Mara Doimo, Timothée Laurent, Andreas Berner, Aldo E Pérez-Rivera, Louise Jenninger, Luis Blanco, Sjoerd Wanrooij, A unique arginine cluster in PolDIP2 enhances nucleotide binding and DNA synthesis by PrimPol., Nucleic acids research, 10.1093/nar/gkab049, 49, 4, 2179-2191, 2021.02.
||Ryo Sugiyama, Kazutoshi Kasho, Kenya Miyoshi, Shogo Ozaki, Wataru Kagawa, Hitoshi Kurumizaka, Tsutomu Katayama, A novel mode of DnaA-DnaA interaction promotes ADP dissociation for reactivation of replication initiation activity., Nucleic acids research, 10.1093/nar/gkz795, 47, 21, 11209-11224, 2019.12.
||Kazutoshi Kasho, Hiroyuki Tanaka, Ryuji Sakai, Tsutomu Katayama, Cooperative DnaA Binding to the Negatively Supercoiled datA Locus Stimulates DnaA-ATP Hydrolysis., The Journal of biological chemistry, 10.1074/jbc.M116.762815, 292, 4, 1251-1266, 2017.01.
||Yukie Inoue, Hiroyuki Tanaka, Kazutoshi Kasho, Taku Oshima, Tsutomu Katayama, Chromosomal location of the DnaA-reactivating sequence DARS2 is important to regulate timely initiation of DNA replication in Escherichia coli, Genes to Cells, 10.1111/gtc.12395, 21, 9, 1015-1023, 2016.09.
||Kazutoshi Kasho, Kazuyuki Fujimitsu, Toshihiro Matoba, Taku Oshima, Tsutomu Katayama, Timely binding of IHF and Fis to DARS2 regulates ATP-DnaA production and replication initiation., Nucleic acids research, 10.1093/nar/gku1051, 42, 21, 13134-49, 2014.12, In Escherichia coli, the ATP-bound form of DnaA (ATP-DnaA) promotes replication initiation. During replication, the bound ATP is hydrolyzed to ADP to yield the ADP-bound form (ADP-DnaA), which is inactive for initiation. The chromosomal site DARS2 facilitates the regeneration of ATP-DnaA by catalyzing nucleotide exchange between free ATP and ADP bound to DnaA. However, the regulatory mechanisms governing this exchange reaction are unclear. Here, using in vitro reconstituted experiments, we show that two nucleoid-associated proteins, IHF and Fis, bind site-specifically to DARS2 to activate coordinately the exchange reaction. The regenerated ATP-DnaA was fully active in replication initiation and underwent DnaA-ATP hydrolysis. ADP-DnaA formed heteromultimeric complexes with IHF and Fis on DARS2, and underwent nucleotide dissociation more efficiently than ATP-DnaA. Consistently, mutant analyses demonstrated that specific binding of IHF and Fis to DARS2 stimulates the formation of ATP-DnaA production, thereby promoting timely initiation. Moreover, we show that IHF-DARS2 binding is temporally regulated during the cell cycle, whereas Fis only binds to DARS2 in exponentially growing cells. These results elucidate the regulation of ATP-DnaA and replication initiation in coordination with the cell cycle and growth phase..
||Kazutoshi Kasho, Tsutomu Katayama, DnaA binding locus datA promotes DnaA-ATP hydrolysis to enable cell cycle-coordinated replication initiation., Proceedings of the National Academy of Sciences of the United States of America, 10.1073/pnas.1212070110, 110, 3, 936-41, 2013.01, The initiation of chromosomal DNA replication is rigidly regulated to ensure that it occurs in a cell cycle-coordinated manner. To ensure this in Escherichia coli, multiple systems regulate the activity of the replication initiator ATP-DnaA. The level of ATP-DnaA increases before initiation after which it drops via DnaA-ATP hydrolysis, yielding initiation-inactive ADP-DnaA. DnaA-ATP hydrolysis is crucial to regulation of initiation and mainly occurs by a replication-coupled feedback mechanism named RIDA (regulatory inactivation of DnaA). Here, we report a second DnaA-ATP hydrolysis system that occurs at the chromosomal site datA. This locus has been annotated as a reservoir for DnaA that binds many DnaA molecules in a manner dependent upon the nucleoid-associated factor IHF (integration host factor), resulting in repression of untimely initiations; however, there is no direct evidence for the binding of many DnaA molecules at this locus. We reveal that a complex consisting of datA and IHF promotes DnaA-ATP hydrolysis in a manner dependent on specific inter-DnaA interactions. Deletion of datA or the ihf gene increased ATP-DnaA levels to the maximal attainable levels in RIDA-defective cells. Cell-cycle analysis suggested that IHF binds to datA just after replication initiation at a time when RIDA is activated. We propose a model in which cell cycle-coordinated ATP-DnaA inactivation is regulated in a concerted manner by RIDA and datA..