The DnaA protein determines the initiation mass of Escherichia coli K-12

Løbner-Olesen, Anders; Skarstad, Kirsten; Hansen, Flemming; Meyenburg, Kaspar von; Boye, Erik

doi:10.1016/0092-8674(89)90802-7

Cited by 303 publications

(272 citation statements)

References 32 publications

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“…Therefore, similar to E. coli DnaA, exchange from ATP-DnaA to ADP-DnaA is crucial for negative regulation for the initiation of chromosome replication in S. aureus. On the other hand, a little increase in origin-to-terminus ratio after induction of wild-type S. aureus DnaA differs from similar studies with E. coli DnaA (7,57), which might be due to differences in initiation control of chromosome replication between them.…”

Section: Inhibition Of Colony Formation and Excessive Initiation By Dcontrasting

confidence: 52%

Rapid Exchange of Bound ADP on the Staphylococcus aureus Replication Initiation Protein DnaA

Kurokawa

Mizumura

Takaki

et al. 2009

Journal of Biological Chemistry

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In Escherichia coli, regulatory inactivation of the replication initiator DnaA occurs after initiation as a result of hydrolysis of bound ATP to ADP, but it has been unknown how DnaA is controlled to coordinate cell growth and chromosomal replication in Gram-positive bacteria such as Staphylococcus aureus. This study examined the roles of ATP binding and its hydrolysis in the regulation of the S. aureus DnaA activity. In vitro, S. aureus DnaA melted S. aureus oriC in the presence of ATP but not ADP by a mechanism independent of ATP hydrolysis. Unlike E. coli DnaA, binding of ADP to S. aureus DnaA was unstable. As a result, at physiological concentrations of ATP, ADP bound to S. aureus DnaA was rapidly exchanged for ATP, thereby regenerating the ability of DnaA to form the open complex in vitro. Therefore, we examined whether formation of ADP-DnaA participates in suppression of replication initiation in vivo. Induction of the R318H mutant of the AAA؉ sensor 2 protein, which has decreased intrinsic ATPase activity, caused over-initiation of chromosome replication in S. aureus, suggesting that formation of ADP-DnaA suppresses the initiation step in S. aureus. Together with the biochemical features of S. aureus DnaA, the weak ability to convert ATP-DnaA into ADP-DnaA and the instability of ADP-DnaA, these results suggest that there may be unidentified system(s) for reducing the cellular ratio of ATPDnaA to ADP-DnaA in S. aureus and thereby delaying the reinitiation of DNA replication.The initiation step of chromosome replication is an important set point for coordinating cell growth and chromosomal replication, and it is tightly regulated so that it takes place only once per cell cycle. DNA replication in bacteria starts when the cell mass reaches an appropriate size and at the appropriate time, which is controlled in part by the quantity and activity of initiator DnaA protein (1-3). In Escherichia coli, DnaA binds to 9-mer DnaA boxes in oriC, the origin of chromosome replication, and melts the duplex at AT-rich 13-mers adjacent to oriC. DnaA then directs loading of the DnaB replicative helicase, which depends on the DnaC helicase loader and priming by the DnaG primase. This is followed by the synthesis of DNA by the DNA polymerase III holoenzyme (4).DnaA is composed of four domains: N-terminal domains I and II are involved in DnaA oligomerization and DnaADnaB interaction, and they show a diversity of sequence among bacteria. Domain III contains AAAϩ motifs, and the C-terminal domain IV mediates DNA binding (3, 5). Biochemical and genetic studies in E. coli have revealed adenine nucleotide binding-mediated control of the DnaA activity (6 -11). Specifically, ATP-bound DnaA actively promotes replication initiation, whereas the ADP-bound form is inert (6, 7). ATP promotes the self-assembly of DnaA at replication origins via a conformational change to form a righthanded helical filament (8, 9). In E. coli, 70% to 80% of DnaA exists as the inactive ADP-bound form, and active ATPDnaA levels increase during the initiation ...

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Section: Inhibition Of Colony Formation and Excessive Initiation By Dcontrasting

confidence: 52%

Rapid Exchange of Bound ADP on the Staphylococcus aureus Replication Initiation Protein DnaA

Kurokawa

Mizumura

Takaki

et al. 2009

Journal of Biological Chemistry

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“…In contrast, gyrB mutations have been described in which the DNA/mass ratio is reduced (Fairweather et al, 1980). In addition, different mutations affecting dnaA have been described which modify the DNA/mass ratio and cause overinitiation at non-permissive temperatures (von Meyenburg and Hansen, 1987), whilst the overexpression of dnaA has been shown to decrease the initiation mass (Lobner-Olesen et al, 1989). However, the present work represents the first clear identification of a cell clock mutant, with an altered initiation mass and an altered cell length under steady-state conditions.…”

Section: Resultsmentioning

confidence: 99%

A mutant cysteinyl-tRNA synthetase affecting timing of chromosomal replication initiation in B. subtilis and conferring resistance to a protein kinase C inhibitor.

et al. 1994

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A Bacilus subdlis mutant spnA95 was isolated as resistant at 30°C to the protein kinase C (PKC) inhibitor, sphinganine, and temperature sensitive for growth. As deduced by flow cytometry measurements, the mutant has a 35% reduced initiation mass at permissive temperature, resulting in initiation of DNA replication much earlier in the cell cycle than in the wild type. This modification is accompanied by a change in cell size, as determined by phase-contrast microscopy and flow cytometry. Therefore, this strain displays the characteristics of a novel cell clock mutant. spnA is a newly identified gene in B.subtilis and was shown to encode a cysteinyl-tRNA synthetase. At non-permissive temperature, the mutant was defective in the synthesis of P70, a protein with several characteristics of PKC (a cysteinerich protein). As one possibility, we propose that the altered timing of replication may be due to the reduced synthesis of specific cysteine-rich proteins normally involved in controlling chromosomal replication initiation in B.subtilis.

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“…The plasmid pALO12 contains the wildtype dnaA gene tagged to the lacP promoter (Lobner-Olesen et al, 1989). The plasmid pTSO200 (Oka et al, 1980) contains the minimal 245 base pair (bp) oriC DNA sequence of E. coli.…”

Section: Methodsmentioning

confidence: 99%

The Bacteriophage λ DNA Replication Protein P Inhibits the oriC DNA- and ATP-binding Functions of the DNA Replication Initiator Protein DnaA of Escherichia coli

Sau¹,

Sil²,

Mandal³

2005

BMB Reports

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Under the condition of expression of λ P protein at lethal level, the oriC DNA-binding activity is significantly affected in wild-type E. coli but not in the rpl mutant. In purified system, the λ P protein inhibits the binding of both oriC DNA and ATP to the wild-type DnaA protein but not to the rpl DnaA protein. We conclude that the λ P protein inhibits the binding of oriC DNA and ATP to the wild-type DnaA protein, which causes the inhibition of host DNA synthesis initiation that ultimately leads to bacterial death. A possible beneficial effect of this interaction of λ P protein with E. coli DNA initiator protein DnaA for phage DNA replication has been proposed.

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The DnaA protein determines the initiation mass of Escherichia coli K-12

Cited by 303 publications

References 32 publications

Rapid Exchange of Bound ADP on the Staphylococcus aureus Replication Initiation Protein DnaA

Rapid Exchange of Bound ADP on the Staphylococcus aureus Replication Initiation Protein DnaA

A mutant cysteinyl-tRNA synthetase affecting timing of chromosomal replication initiation in B. subtilis and conferring resistance to a protein kinase C inhibitor.

The Bacteriophage λ DNA Replication Protein P Inhibits the oriC DNA- and ATP-binding Functions of the DNA Replication Initiator Protein DnaA of Escherichia coli

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