Studies on the Folded Chromosome of Escherichia coli

Worcel, Abraham; Burgi, Elizabeth; Robinton, J.; Carlson, C.

doi:10.1101/sqb.1974.038.01.007

Cited by 40 publications

(22 citation statements)

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“…DNA release does not occur even in 1% Sarcosyl at 37 C, but does so at60Cin4MNaCl (16,17), orat70Cin0.15 M NaCl, or by phenol treatment at pH 8 at room temperature (unpublished data), and the liberated DNA is in the denatured form. The properties of these complexes are reminiscent of cell envelope proteins relatively insensitive to proteolytic enzymes and detergents (5,32). This is in accord with the binding of transforming DNA by B. subtilis cell membranes, revealed by means of fractionation of cell lysates in a Renografin gradient (3) and by autoradiography (30).…”

Section: -mentioning

confidence: 85%

“…In view of the interest in DNA-protein complexes (2a, 3, 4, 16), it is worth noting that: (i) no such complex is found in a system containing native or denatured DNA plus all other constituents except cells (16); (ii) the properties of the complex are reminiscent of complexes with membrane proteins (5,32); (iii) the specificity of complex formation depends on the nature of the DNA, as shown in this study. This points to involvement in the complex of a cellular constituent(s) and not of added proteins such as lysozyme or Pronase (2a).…”

Section: -mentioning

confidence: 99%

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Heterologous deoxyribonucleic acid uptake and complexing with cellular constituents in competent Bacillus subtilis

Soltyk¹,

Shugar²,

Piechowska³

1975

J Bacteriol

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With competent cultures of Bacillus subtilis the uptake of Escherichia coli deoxyribonucleic acid (DNA) is about 50% that for homologous DNA. Uptake of phage T6 DNA, if any, is of the order of 7%, while nonglucosylated phage T6 (T*6) DNA is taken up almost as effectively as homologous DNA. Both T6 and T4 richia coli 15T-and phages T4 and T6, provided by Irena Pietrzykowska of this Institute. The phages were cultivated on E. coli BB, obtained from W. Szybalski of the University of Wisconsin. Nonglucosylated phage T6 DNA (T*6 DNA) was prepared from phage T6 cultivated on the uridine diphosphoglucose phosphorylase-defective mutant of E. coli B-4°Luria no. 56 (6), provided by S. E. Luria of the Massachusetts Institute of Technology. The nonglucosylated phage T6 (T*6) titer was determined with the aid of the strain Shigella dysenteriae, supplied by N. Symonds of the University of Sussex, England. Absence of glucosylation was confirmed by 1429 on August 3, 2020 by guest

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Section: -mentioning

confidence: 85%

Section: -mentioning

confidence: 99%

Heterologous deoxyribonucleic acid uptake and complexing with cellular constituents in competent Bacillus subtilis

Soltyk¹,

Shugar²,

Piechowska³

1975

J Bacteriol

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“…Effects of m.o.e, on the nueleoid structure DNA in uninfected E. eoli has a condensed and folded tertiary structure (Stonington & Pettijohn, 1971;Pettijohn & Hecht, 1973;Worcel et al, 1973). Nucleoids containing phage DNA are present after infection with T4 or T7 (Hamilton & Pettijohn, 1976;Helland, 1977).…”

Section: Inhibition Oft7 Development 121mentioning

confidence: 99%

Inhibition of T7 Development at High Concentrations of the Phage

Haarr

Jensen

Helland

1981

Journal of General Virology

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SUMMARYEscherichia coli B exposed to high doses of bacteriophage T7 did not lyse. A similar effect was observed when the high dose was added within the first 7 rain after primary infection. No viable phage was formed. DNA synthesis was inhibited rapidly and the nucleoid structure was'absent. Protein synthesis was in general markedly reduced and so were the activities of the phage-specific enzymes endolysin and DNA polymerase. However, phage genes were transcribed both by the host RNA polymerase and by the phage-specific enzyme. We suggest that inhibition of phage development is due to structural alterations occurring in the cell wall/membrane such that replication is inhibited.

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“…Jacob et al (18) proposed that DNA strand segregation may occur via specific DNA-membrane attachments, a view indirectly supported by observations that the replication origin, terminus, and fork are membrane-associated in vi-tro (21). Since chromosomes associated with large fragments of the cell envelope can be isolated (20,32,44,45), both DNA packaging and membrane association can be studied in a single system.…”

mentioning

confidence: 99%

“…Membrane-associated chromosomes can be isolated either in high salt (32,45), or in low salt plus spermidine (20). Although the high-salt and spermidine chromosomes are similar, the present study focuses on those prepared in spermidine because they should better reflect in vivo DNA-membrane interactions; they contain more proteins (33) and are able to undergo replicative-type DNA synthesis (20).…”

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confidence: 99%

Association of the Folded Chromosome with the Cell Envelope of Escherichia coli : Nature of the Membrane-Associated DNA

1978

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Membrane-associated folded chromosomes isolated from Escherichia coli in the presence of spermidine sedimented at about 5,800S. The folded chromosome and the membrane fragment were each stable in .the absence of the other; a 1,700S folded chromosome was obtained after removal of the membrane by a Sarkosyl treatment, and a 4,OOOS membrane fragment remained after digestion of the chromosomal DNA with deoxyribonuclease I. The interaction between the folded chromosome and the membrane fragment was stable, and, even when the DNA was unfolded, both components remained associated and cosedimented. The large frictional effect of the unfolded DNA reduced the sedimentation rate of the complex to about 2,000S. Partial removal of this unfolded DNA with restriction endonucleases caused the membrane fragments and the remaining associated DNA to sediment faster, at about 3,500S. The DNA remaining associated with the membrane fragments after restriction endonuclease treatment, about 4.5% of the total DNA when EcoRI was used, was indistinguishable from the DNA released from the membranes by three criteria: (i) DNA size distribution in agarose gels after electrophoresis, (ii) reassociation kinetics, and (iii) thermal elution from hydroxylapatite. This finding, that random DNA sequences rather than specific ones were responsible for the majority of the DNA-membrane interactions, argues against the folded chromosome's being a static structure with specific DNA sequences interacting with the cell envelope. t Present address:

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Studies on the Folded Chromosome of Escherichia coli

Cited by 40 publications

References 0 publications

Heterologous deoxyribonucleic acid uptake and complexing with cellular constituents in competent Bacillus subtilis

Heterologous deoxyribonucleic acid uptake and complexing with cellular constituents in competent Bacillus subtilis

Inhibition of T7 Development at High Concentrations of the Phage

Association of the Folded Chromosome with the Cell Envelope of Escherichia coli : Nature of the Membrane-Associated DNA

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