Isolation and Characterization of Thermosensitive
            <i>Escherichia coli</i>
            Mutants Defective in Deoxyribonucleic Acid Replication

Wechsler, James A.; Nüsslein, Volker; Otto, Bernd; Klein, Albrecht; Bonhoeffer, Friedrich; Herrmann, Richard; Gloger, Lieselotte; Schaller, Heinz

doi:10.1128/jb.113.3.1381-1388.1973

Cited by 101 publications

(28 citation statements)

References 6 publications

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“…A heat sensitive defect caused by a single mutation in the uptake or synthesis of a common precursor, needed for both DNA and RNA synthesis (FUCHS et al 1972, FUCHS andNEUHARD 1973) could hardly explain our observation of an immediate stop of both syntheses (Fig. 3A, B, C), because the cellular pool of precursors should allow a limited continuation of syntheses (WECHSLER et al 1973, Born et al 1981, BLINKOVA et al 1983). In addition, in our pulse labelling experiments (Fig.…”

Section: Discussionmentioning

confidence: 77%

Characterization of a thermosensitive mutant of Streptomyces hygroscopicus defective in both DNA and RNA syntheses

Süss¹,

Frunder²,

Klaus³

et al. 1988

J. Basic Microbiol.

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A stable temperature sensitive mutant of Streptomyces hygroscopicus JA6599 defective in both DNA and RNA syntheses is described. The mutant ts35 is characterized by an immediate stop of DNA synthesis and continued protein synthesis after transfer to restrictive temperature. The reinitiation of DNA synthesis begins immediately after a return to the permissive temperature. This kinetics of macromolecular synthesis at restrictive temperature appears to share similarities with a defect in the DNA elongation process, as described for Escherichia coli (Carl 1970, Hanna and Carl 1975). The simultaneous stop of both DNA and RNA syntheses may be caused by an additional mutational event affecting also the RNA synthesis. The data were discussed with respect to similar results in E. coli.

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Section: Discussionmentioning

confidence: 77%

Characterization of a thermosensitive mutant of Streptomyces hygroscopicus defective in both DNA and RNA syntheses

Süss¹,

Frunder²,

Klaus³

et al. 1988

J. Basic Microbiol.

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“…The Ecoli K-12 strains used were: C600 (Bachmann, 1972), CC118 (Manoil and Beckwith, 1985), CSH50 (Miller, 1972) and BTIOOO (Wechsler et al, 1973). PAO1024 (r-, m+; from K.Nordstrom) was the Paeruginosa strain used.…”

Section: Bacterial Strainsmentioning

confidence: 99%

A leucine zipper motif determines different functions in a DNA replication protein.

et al. 1996

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RepA is the replication initiator protein of the Pseudomonas plasmid pPS10 and is also able to autoregulate its own synthesis. Here we report a genetic and functional analysis of a leucine zipper‐like (LZ) motif located at the N‐terminus of RepA. It is shown that the LZ motif modulates the equilibrium between monomeric and dimeric forms of the protein and that monomers of RepA interact with sequences at the origin of replication, oriV, while dimers are required for interactions of RepA at the repA promoter. Further, different residues of the LZ motif are seen to have different functional roles. Leucines at the d positions of the putative alpha‐helix are relevant in the formation of RepA dimers required for transcriptional autoregulation. They also modulate other RepA‐RepA interactions that result in cooperative binding of protein monomers to the origin of replication. The residues at the b/f positions of the putative helix play no relevant role in RepA‐RepA interactions. These residues do not affect RepA autoregulation but do influence replication, as demonstrated by mutants that, without affecting binding to oriV, either increase the host range of the plasmid or are inactive in replication. It is proposed that residues in b/f positions play a relevant role in interactions between RepA and host replication factors.

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“…The Escherichia coli K‐12 polA + strain C600 [1] and the polA − strain BT1000 [17] were used to prepare plasmid‐free extracts. SSC1 (pMS4D1) overproducing Gpα[16] was used to prepare extracts enriched in Gpα.…”

Section: Methodsmentioning

confidence: 99%

Enhancement of ColE1-type replication promoted by the Gpα initiator protein of bacteriophage P4

Dıćaz-Orejas

Rückert

2006

FEMS Microbiology Letters

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Gp alpha, the phage P4 specific replication protein, increases in vitro replication of pCN51, a pBR322 based replicon, by a factor of two. This effect is dependent on DNA polymerase I and requires transcription by host RNA polymerase. Electron microscopic analysis of replicating intermediates indicates that pCN51 replication occurred from the same origin and with the same directionality in the presence and in the absence of Gp alpha. These results reveal that Gp alpha can influence the replication of an heterologous replicon and show that this effect occurs in ColE1-type replicons without altering the normal pattern of initiation. Further analysis of replicating intermediates shows an increase in the average size of the ColE1-type replication 'bubble' obtained in the presence of Gp alpha. It is proposed that Gp alpha interacts with the ColE1 replisome complex at an early replication stage.

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Isolation and Characterization of Thermosensitive Escherichia coli Mutants Defective in Deoxyribonucleic Acid Replication

Abstract: Media. Tris(hydroxymethyl )aminomethane (Tris)-minimal medium contains (per liter): 0.5 g of NaCl, 8.0 g of KCI, 12.2 g of Tris, 1.2 g of NH4C1, 0.8 g of sodium pyruvate, 0.2 g of MgC1,, 0.5 g of (NH4)2-SO4 10H2O, 0.01 g of thiamine, 2 g of glucose, 0.44 g 1381 on July 16, 2020 by guest

Cited by 101 publications

References 6 publications

Characterization of a thermosensitive mutant of Streptomyces hygroscopicus defective in both DNA and RNA syntheses

Characterization of a thermosensitive mutant of Streptomyces hygroscopicus defective in both DNA and RNA syntheses

A leucine zipper motif determines different functions in a DNA replication protein.

Enhancement of ColE1-type replication promoted by the Gpα initiator protein of bacteriophage P4

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