Negative control of DNA replication by hydrolysis of ATP bound to DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli

DnaA protein, the initiation factor for chromosomal DNA replication in Escherichia coli, is activated by ATP. ATP bound to DnaA protein is slowly hydrolyzed to ADP, but the physiological role of ATP hydrolysis is unclear. We constructed, by site‐directed mutagenesis, mutated DnaA protein with lower ATPase activity, and we examined its function in vitro and in vivo. The ATPase activity of purified mutated DnaA protein (Glu204→Gln) decreased to one‐third that of the wild‐type DnaA protein. The mutation did not significantly affect the affinity of DnaA protein for ATP or ADP. The mutant dnaA gene showed lethality in wild‐type cells but not in cells growing independently of the function of oriC. Induction of the mutated DnaA protein in wild‐type cells caused an overinitiation of DNA replication. Our results lead to the thesis that the intrinsic ATPase activity of DnaA protein negatively regulates chromosomal DNA replication in E.coli cells.

[1]  T. Katayama,et al.  Conformational transition of DnaA protein by ATP: structural analysis of DnaA protein, the initiator of Escherichia coli chromosome replication. , 1997, Biochemical and biophysical research communications.

[2]  T. Katayama,et al.  Molecular Design of Inhibitors of in Vitro oriC DNA Replication Based on the Potential to Block the ATP Binding of DnaA Protein* , 1996, The Journal of Biological Chemistry.

[3]  T. Katayama,et al.  Effect on DNA topology by DnaA protein, the initiation factor of chromosomal DNA replication in Escherichia coli. , 1996, Biochemistry.

[4]  Masasuke Yoshida,et al.  Characterization of a Potential Catalytic Residue, Asp-133, in the High Affinity ATP-binding Site of Escherichia coli SecA, Translocation ATPase* , 1996, The Journal of Biological Chemistry.

[5]  Y. Ishikawa,et al.  Influence of Cluster Formation of Acidic Phospholipids on Decrease in the Affinity for ATP of DnaA Protein (*) , 1996, The Journal of Biological Chemistry.

[6]  T. Katayama,et al.  Characterization of Escherichia coli DnaAcos protein in replication systems reconstituted with highly purified proteins , 1995, Molecular microbiology.

[7]  T. Katayama,et al.  DnaA Protein Is Sensitive to a Soluble Factor and Is Specifically Inactivated for Initiation of in Vitro Replication of the Escherichia coli Minichromosome (*) , 1995, The Journal of Biological Chemistry.

[8]  Masasuke Yoshida,et al.  A common topology of proteins catalyzing ATP‐triggered reactions , 1995, FEBS letters.

[9]  W. Dowhan,et al.  In vivo evidence for the involvement of anionic phospholipids in initiation of DNA replication in Escherichia coli. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[10]  T. Katayama.  The mutant DnaAcos protein which overinitiates replication of the Escherichia coli chromosome is inert to negative regulation for initiation. , 1994, The Journal of biological chemistry.

[11]  Masasuke Yoshida,et al.  Spatial precision of a catalytic carboxylate of F1‐ATPase β subunit probed by introducing different carboxylate‐containing side chains , 1994, FEBS letters.

[12]  A. Kornberg,et al.  Hyperactive initiation of chromosomal replication in vivo and in vitro by a mutant initiator protein, DnaAcos, of Escherichia coli. , 1994, The Journal of biological chemistry.

[13]  A. Kornberg,et al.  Fluid membranes with acidic domains activate DnaA, the initiator protein of replication in Escherichia coli. , 1993, The Journal of biological chemistry.

[14]  E. Kimura,et al.  Complementation of two overlapping fragments of SecA, a protein translocation ATPase of Escherichia coli, allows ATP binding to its amino-terminal region. , 1990, The Journal of biological chemistry.

[15]  M. Akita,et al.  SecA interacts with secretory proteins by recognizing the positive charge at the amino terminus of the signal peptide in Escherichia coli. , 1990, The Journal of biological chemistry.

[16]  Frank McCormick,et al.  ras GTPase activating protein: Signal transmitter and signal terminator , 1989, Cell.

[17]  A. Kornberg,et al.  Membrane attachment activates dnaA protein, the initiation protein of chromosome replication in Escherichia coli. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[18]  A. Kornberg,et al.  The dnaA protein of Escherichia coli. Abundance, improved purification, and membrane binding. , 1988, The Journal of biological chemistry.

[19]  A. Kornberg,et al.  Sequential early stages in the in vitro initiation of replication at the origin of the Escherichia coli chromosome. , 1988, The Journal of biological chemistry.

[20]  A Kornberg,et al.  Cardiolipin activation of dnaA protein, the initiation protein of replication in Escherichia coli. , 1988, The Journal of biological chemistry.

[21]  A. Kornberg,et al.  Duplex opening by dnaA protein at novel sequences in initiation of replication at the origin of the E. coli chromosome , 1988, Cell.

[22]  Masasuke Yoshida,et al.  Invitro mutated β subunits from the F1-ATPase of the thermophilic bacterium, PS3, containing glutamine in place of glutamic acid in positions 190 or 201 assembles with the α and γ subunits to produce inactive complexes , 1987 .

[23]  A. Kornberg,et al.  ATP activates dnaA protein in initiating replication of plasmids bearing the origin of the E. coli chromosome , 1987, Cell.

[24]  A. Gilman,et al.  G proteins: transducers of receptor-generated signals. , 1987, Annual review of biochemistry.

[25]  Thomas A. Kunkel,et al.  Rapid and efficient site-specific mutagenesis without phenotypic selection. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Arthur Kornberg,et al.  The dnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites , 1984, Cell.

[27]  A. Kornberg,et al.  Purified dnaA protein in initiation of replication at the Escherichia coli chromosomal origin of replication. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[28]  A. Kornberg,et al.  Enzymatic replication of the origin of the Escherichia coli chromosome. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[29]  M. Gefter,et al.  DNA Replication , 2019, Advances in Experimental Medicine and Biology.

[30]  F. Jacob,et al.  On the process of cellular division in Escherichia coli. 3. Thermosensitive mutants of Escherichia coli altered in the process of DNA initiation. , 1970, Journal of molecular biology.

[31]  K. Lark,et al.  Initiation of DNA replication in Escherichia coli 15T-: chronological dissection of three physiological processes required for initiation. , 1969, Journal of molecular biology.