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A new mutant of Bacillus subtilis altered in the initiation of chromosome replication

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Summary

We have isolated a new mutant of Bacillus subtilis temperature sensitive in DNA replication; its properties are those of an initiation mutant. When liquid cultures are shifted to 48° DNA replication is the first macromolecular synthesis that stops, but only after synthesis of the amount of DNA predicted for the completion of one replication round. When spores of the mutant are germinated and shifted to 48° at subsequent times, one round of DNA replication is observed only when the shift occurs between 60 and 100 min; earlier shifts do not allow replication to start, later shifts allow more than one replication. The DNA replicated after a shift to high temperature is enriched in markers close to the terminus. The reinitiation of DNA replication stopped by the high temperature, takes place following a shift to a permissive temperature only if protein synthesis is allowed. Examination of DNA replication following toluene treatment shows that the elongation of DNA chains is not affected at the non-permissive temperature.

This mutant is shown by PBS-1 mapping to correspond to a new gene denominated dna P, which is located between the thy A and fur A genes and is distinct from all the mapped dna and rec genes of Bacillus subtilis. The mutation confers to the cells also a deficiency in the ability to be transformed, to be transfected with SPP1 phage DNA, and to survive treatment with methyl-methane sulfonate. These deficiencies, observed at the permissive temperature, are no more temperature dependent than in the parental strain. The ability to perform homologous and heterologous transduction with PBS-1 phage and the sensitivity to ultraviolet radiation or mitomycin C are normal.

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Abbreviations

CAP:

chloramphenicol

EDTA:

sodium ethylene-diamino-tetraacetate

HPura:

6-(p-hydroxyphenylazo)-uracil

KPB:

potassium phosphate buffer, 0.05 M, pH 7.4

MMS:

methylmethane-sulfonate

NTG:

N-methyl-N-nitro-N-nitrosoguanidine

SSC:

standard saline citrate solution (0.15 M NaCl, 0.015 M sodium citrate)

TCA:

trichloroacetic acid

Tris:

tris-hydroxymethyl-aminomethane

UV:

ultraviolet light

mic :

minimum inhibitory concentration

pfu :

plaque forming unit

moi :

multiplicity of infection

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Authors and Affiliations

  1. Laboratorio di Genetica, Biochemica ed Evoluzionistica del Consiglio Nazionale delle Ricerche, Università di Pavia, Pavia, Italy

    Silvano Riva, Cornelis van Sluis, Giorgio Mastromei, Carmen Attolini, Giorgio Mazza, Mario Polsinelli & Arturo Falaschi

  2. Istituto di Genetica, Università di Pavia, Pavia, Italy

    Silvano Riva, Giorgio Mastromei, Carmen Attolini, Giorgio Mazza, Mario Polsinelli & Arturo Falaschi

  3. Department of Biochemistry, Leiden State University, Leiden, The Netherlands

    Cornelis van Sluis

  4. Cattedra di Genetica, Università di Firenze, Via Romana 17, I-50100, Firenze, Italy

    Mario Polsinelli

Authors
  1. Silvano Riva
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  2. Cornelis van Sluis
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  3. Giorgio Mastromei
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  4. Carmen Attolini
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  5. Giorgio Mazza
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  6. Mario Polsinelli
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  7. Arturo Falaschi
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Communicated by F. Gros

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Riva, S., van Sluis, C., Mastromei, G. et al. A new mutant of Bacillus subtilis altered in the initiation of chromosome replication. Molec. gen. Genet. 137, 185–202 (1975). https://doi.org/10.1007/BF00333015

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  • DOI: https://doi.org/10.1007/BF00333015

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