DnaA initiator—also a transcription factor

Messer, Walter; Weigel, Christoph

doi:10.1046/j.1365-2958.1997.3171678.x

Cited by 146 publications

(136 citation statements)

References 57 publications

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“…Another explanation for the effect of DnaA (or datA) on cell division could be that DnaA, being a transcription factor, affects transcription of cell division genes (Messer & Weigel, 1997). There are several putative DnaA boxes in the ftsQAZ operons (Robinson et al, 1984), but DnaA control of fts expression has been controversial and different experimental setups have given different results (Garrido et al, 1993;Masters et al, 1989;Smith et al, 1996).…”

Section: The Inhibitory Effect Of Data Is Not Dependent On the Sos Rementioning

confidence: 99%

The Escherichia coli datA site promotes proper regulation of cell division

Flåtten

Skarstad

2014

Microbiology

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In Escherichia coli inhibition of replication leads to a block of cell division. This checkpoint mechanism ensures that no cell divides without having two complete copies of the genome to pass on to the two daughter cells. The chromosomal datA site is a 1 kb region that contains binding sites for the DnaA replication initiator protein, and which contributes to the inactivation of DnaA. An excess of datA sites provided on plasmids has been found to lead to both a delay in initiation of replication and in cell division during exponential growth. Here we have investigated the effect of datA on the cell division block that occurs upon inhibition of replication initiation in a dnaC2 mutant. We found that this checkpoint mechanism was aided by the presence of datA. In cells where datA was deleted or an excess of DnaA was provided, cell division occurred in the absence of replication and anucleate cells were formed. This finding indicates that loss of datA and/or excess of DnaA protein promote cell division. This conclusion was supported by the finding that the lethality of the division-compromised mutants ftsZ84 and ftsI23 was suppressed by deletion of datA, at the lowest non-permissive temperature. We propose that the cell division block that occurs upon inhibition of DNA replication is, at least in part, due to a drop in the concentration of the ATP-DnaA protein. INTRODUCTIONIn Escherichia coli cell division involves the formation of a septum that separates the cell into two daughter cells. It is crucial that the two daughter nucleoids are segregated into each half of the cell, so that each daughter cell receives a copy of the genetic material and that none of the nucleoids are trapped in the septum. To ensure this, cell division and chromosome segregation are highly regulated processes involving a number of different proteins (see Egan & Vollmer, 2013;Harry et al., 2006;and Weiss, 2004, for reviews), the most studied being FtsZ. This protein polymerizes to form a ring-like structure at the site of division and serves as a scaffold for recruitment of other division proteins (Bi & Lutkenhaus, 1991). How the timing of the formation of the FtsZ ring is regulated is not clear.It is known that the chromosome replication cycle is independent of the cell division cycle, but that cell division is dependent on chromosome replication (Boye & Nordström, 2003; Nordström et al., 1991). That is, a blockage of chromosome replication stops cell division (Helmstetter & Pierucci, 1968;Walker & Pardee, 1968). Thus, a checkpoint mechanism must exist. The mechanism ensures that cell division does not occur in the absence of replication and thus prevents the formation of DNA-free (anucleate) daughter cells (Boye & Nordström, 2003; Nordström et al., 1991). Whether this mechanism is related to other regulatory circuits necessary for coupling the division cycle to cell physiology and to the growth rate of the cells, is not clear (see Haeusser & Levin, 2008, for a review).The DnaA protein is one of the main players in initiation of replica...

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Section: The Inhibitory Effect Of Data Is Not Dependent On the Sos Rementioning

confidence: 99%

The Escherichia coli datA site promotes proper regulation of cell division

Flåtten

Skarstad

2014

Microbiology

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“…These data suggest that disruptants maintained a low level of metabolic activity during the stationary phase to preserve cellular energy and enhance viability. DnaA could also directly regulate expression of these genes, given its function as a transcription factor in many bacteria (Messer and Weigel, 1997;Ishikawa et al, 2007;McAdams and Shapiro, 2009). …”

Section: Increased Viability Of Dnaa Disruptants Under Long Culture Cmentioning

confidence: 99%

Diversification of DnaA dependency for DNA replication in cyanobacterial evolution

et al. 2015

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Regulating DNA replication is essential for all living cells. The DNA replication initiation factor DnaA is highly conserved in prokaryotes and is required for accurate initiation of chromosomal replication at oriC. DnaA-independent free-living bacteria have not been identified. The dnaA gene is absent in plastids and some symbiotic bacteria, although it is not known when or how DnaA-independent mechanisms were acquired. Here, we show that the degree of dependency of DNA replication on DnaA varies among cyanobacterial species. Deletion of the dnaA gene in Synechococcus elongatus PCC 7942 shifted DNA replication from oriC to a different site as a result of the integration of an episomal plasmid. Moreover, viability during the stationary phase was higher in dnaA disruptants than in wild-type cells. Deletion of dnaA did not affect DNA replication or cell growth in Synechocystis sp. PCC 6803 or Anabaena sp. PCC 7120, indicating that functional dependency on DnaA was already lost in some nonsymbiotic cyanobacterial lineages during diversification. Therefore, we proposed that cyanobacteria acquired DnaA-independent replication mechanisms before symbiosis and such an ancestral cyanobacterium was the sole primary endosymbiont to form a plastid precursor.

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“…The DnaA protein recognizes and binds speciWcally to consensus sequences in oriC, termed DnaA boxes (R1-R4 and M) [13]. In addition to its central role in the initiation of replication, DnaA acts as a transcription factor of various genes [14]. On binding to DnaA box motifs located within promoters or coding regions of these genes, DnaA can act as a repressor, an activator, or a terminator of transcription [15].…”

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confidence: 99%

Protein microarray technology and ultraviolet crosslinking combined with mass spectrometry for the analysis of protein–DNA interactions

Kersten

Possling

Blaesing

et al. 2004

Analytical Biochemistry

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To gain insights into complex biological processes, such as transcription and replication, the analysis of protein-DNA interactions and the determination of their sequence requirements are of central importance. In this study, we probed protein microarray technology and ultraviolet crosslinking combined with mass spectrometry (MS) for their practicability to study protein-DNA interactions. We chose as a model system the well-characterized interaction of bacterial replication initiator DnaA with its cognate binding site, the DnaA box. Interactions of DnaA domain 4 with a high-aYnity DnaA box (R4) and with a low-aYnity DnaA box (R3) were compared. A mutant DnaA domain 4, A440V, was included in the study. DnaA domain 4, wt, spotted onto FAST slides, revealed a strong signal only with a Cy5-labeled, double-stranded, 21-mer oligonucleotide containing DnaA box R4. No signals were obtained when applying the mutant protein. Ultraviolet crosslinking combined with nanoLC/MALDI-TOF MS located the site of interaction to a peptide spanning amino acids 433-442 of Escherichia coli DnaA. This fragment contains six residues that were identiWed as being involved in DNA binding by recently published crystal structure and nuclear magnetic resonance (NMR) analysis. In the future, the technologies applied in this study will become important tools for studying protein-DNA interactions.  2004 Elsevier Inc. All rights reserved.

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DnaA initiator—also a transcription factor

Cited by 146 publications

References 57 publications

The Escherichia coli datA site promotes proper regulation of cell division

The Escherichia coli datA site promotes proper regulation of cell division

Diversification of DnaA dependency for DNA replication in cyanobacterial evolution

Protein microarray technology and ultraviolet crosslinking combined with mass spectrometry for the analysis of protein–DNA interactions

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