Control of Deoxyribonucleic Acid Synthesis in a
            <i>Bacillus subtilis</i>
            Mutant Temperature Sensitive for Initiation of Chromosome Replication

Laurent, Simone J.

doi:10.1128/jb.117.2.329-336.1974

Cited by 10 publications

(4 citation statements)

References 19 publications

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“…Furthermore, the cells retain complete viability during the period for which they were at 45 C. As in the case of B. subtilis 168 ts 37, an initiation mutant isolated by Laurent and Vannier (7)(8)(9), dna-20(Ts) has been found particularly useful in preliminary studies on possible transcriptional and translational events associated with the initiation of chromosome replication (P. Upcroft, H. J. Dyson, and R. G. Wake, Proc. Aust.…”

Section: Discussionmentioning

confidence: 99%

Characteristics of a Bacillus subtilis W23 mutant temperature sensitive for initiation of chromosome replication

Upcroft¹,

Dyson²,

Wake³

1975

J Bacteriol

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A temperature-sensitive mutant of Bacillus subtilis W23, dna-20 (Ts), has been isolated and shown to be defective in initiation of rounds of chromosome replication at the nonpermissive temperature. Upon transfer of dna-20(Ts) from 30 to 45 C, deoxyribonucleic acid synthesis, as measured by [3H]thymine incorporation, gradually ceases. The distribution of genetic markers among unreplicated and replicated deoxyribonucleic acid, isolated from dna-20(Ts) after a period at 43 C in a medium containing 5-bromouracil, and fractionated in a CsCl gradient, shows that the cessation of initiation at the higher temperature is immediate. On the other hand, ribonucleic acid and protein synthesis continues at elevated or unaltered rates for some time after the shift to 45 C. Marker frequency analysis shows that all rounds of replication in progress at the time of the temperature shift terminate rapidly (within 40 min), even when chromosomes are replicating dichotomously in rich media. dna-20(Ts) remains 100% viable for at least 2 h at 45 C. Over a 5-h period at 45 C the nuclear bodies remain compact; a small number (less than 5%) of deoxyribonucleic acid-less cells are produced, but there is no morphological distortion of the cells. When the cells are returned to 30 C after 2 h at 45 C, chromosome replication is initiated rapidly at the normal origin and then proceeds in the normal established sequence. However, a second round of replication is initiated soon after the first. dna-20(Ts) has been shown to map as a B-group mutation, the major class of initiation mutants identified in B. subtillus 168.

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Section: Discussionmentioning

confidence: 99%

Characteristics of a Bacillus subtilis W23 mutant temperature sensitive for initiation of chromosome replication

Upcroft¹,

Dyson²,

Wake³

1975

J Bacteriol

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“…Initiation of the first cycle requires RNA synthesis but not protein synthesis (17). Depending on the length of the preceding period at a nonpermissive temperature, the initiation of the second cycle becomes chloramphenicol resistant and rifampin resistant during the first cycle, well before the second cycle itself begins (18). This paper shows that, after purification and complete denaturation of DNA, an in vivo-labeled RNA is found associated with high-molecular-weight DNA.…”

mentioning

confidence: 81%

“…wave showed a maximum between 30 and 40 min, i.e., during the elongation of the first replication cycle and thus before the initiation of the second cycle. In an earlier study (18), it was shown that, after a period at 45C, the initiation of the second cycle became chloramphenicol resistant and rifampin resistant before the initia-tion itself began. (iii) We have not yet studied the time of appearance of the initiation potential for the third replication cycle.…”

Section: Methodsmentioning

confidence: 99%

“…Our earlier studies have shown that these proteins appear in the bacteria before the initiation itself. Moreover, the longer the preceding period at 45°C, the earlier the bacteria acquire initiation proteins for the second round (18). The effect of the length of the period at 45°C can be explained by a model involving two categories of processes.…”

Section: Methodsmentioning

confidence: 99%

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Possible involvement of DNA-linked RNA in the initiation of Bacillus subtilis chromosome replication

Henckes¹,

Vannier²,

Buu³

et al. 1982

J Bacteriol

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After thermal denaturation, an in vivo-labeled RNA was found in a temperature-sensitive initiation mutant of Bacillus subtilis (dna-37) associated with highmolecular-weight DNA. This RNA could be clearly distinguished from other RNA species by different techniques of separation, such as Sepharose 2B filtration, chromatography on nitrocellulose, and equilibrium centrifugation in density gradients. It was obtained even when HCHO was present during denaturation and chilling of nucleic acids and was still detected after a second denaturation as well as after incubation with proteinase K. Properties of the complex were not altered by prior treatment with RNase H. A control .experiment using two samples of the complex treated either with pancreatic DNase or with pancreatic RNase, denatured together and centrifuged in the same density gradient, showed that no artifactual associations occur between the DNA and the RNA components of the complex. These results demonstrate that the DNA and RNA in the complex are associated by neither hydrogen bonds nor proteins, but are indicative of a DNA-RNA covalent linkage. In addition, during synchronous replication after a previous period at a nonpermissive temperature, DNA-linked RNA synthesis took place at specific times which coincided with the appearance of rifampin resistance of the first and the second replication cycles. A possible involvement of this RNA in the initiation of chromosome replication is discussed.

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Molecular cloning of Bacillus subtilis genes involved in DNA metabolism

et al. 1987

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Different clones carrying a chromosomal DNA fragment able to transform Bacillus subtilis mutants dnaA13, dnaB19, dnaG5, recG40 and polA42 to a wild-type phenotype were isolated from a library constructed in plasmid pJH101. A lambda recombinant clone carrying a chromosomal fragment able to transform dnaC mutants was obtained from a lambda Charon 4A library. A restriction map of the cloned DNA fragments was constructed. The 11.3 kb cloned DNA fragment of plasmid pMP60-13 containing the wild-type sequence of dnaG5 was shown to transform a recF33 mutant as well.

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Control of Deoxyribonucleic Acid Synthesis in a Bacillus subtilis Mutant Temperature Sensitive for Initiation of Chromosome Replication

Cited by 10 publications

References 19 publications

Characteristics of a Bacillus subtilis W23 mutant temperature sensitive for initiation of chromosome replication

Characteristics of a Bacillus subtilis W23 mutant temperature sensitive for initiation of chromosome replication

Possible involvement of DNA-linked RNA in the initiation of Bacillus subtilis chromosome replication

Molecular cloning of Bacillus subtilis genes involved in DNA metabolism

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