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PubMed1982

Plasmids from Staphylococcus aureus replicate in yeast Saccharomyces cerevisiae.

It is known that some plasmids, such as RP4, can replicate in many Gram-negative bacteria. Certain small Staphylococcus aureus plasmids have an even broader host range, being able to replicate in not only phylogenetically distant Gram-positive bacteria such as Bacillus subtilis or Streptococcus pneumoniae, but also in the Gram-negative bacterium Escherichia coli. Here we have examined whether these plasmids can also replicate in a lower eukaryote, the yeast Saccharomyces cerevisiae. For this purpose we constructed hybrids between a S. aureus plasmid pC194 and an E. coli plasmid YIp5, which carries a ura-3 gene easy to select for in yeast but cannot replicate in this host. We found that the hybrids transformed yeast with high efficiency (as did hybrids between YIp5 and three other S. aureus plasmids); were maintained extrachromosomally in yeast; and were not modified during residence in yeast. We conclude from this evidence that S. aureus plasmids can replicate in yeast, which raises the questions of whether the replication signals used by prokaryotes and eukaryotes are similar, and how far up the phylogenetic tree the organisms still able to be hosts to S. aureus plasmids may be.

Base Sequence↗
PubMedDate not supplied

Role of reverse transcription in the generation of extrachromosomal copia mobile genetic elements.

The Drosophila genetic element copia is one of the best studied eukaryotic transposable sequences. Copia shares structural features with a wide variety of mobile elements in Drosophila, Lepidoptera, yeast and vertebrates, the last class being the retrovirus proviruses. Furthermore, retrovirus-like particles containing copia RNA have been isolated from Drosophila cells and extrachromosomal circular copias with structures closely resembling circular retrovirus proviruses have been isolated and cloned. Therefore, copia-like elements and retroviruses may be members of a class of mobile genetic elements existing throughout the eukaryotic kingdom. Consequently, there has been speculation that retroviruses evolved from transposable elements and, conversely, that copia-like elements transpose as retroviruses or retrovirus-like particles. To date, however, there has been no demonstration that copia RNA is reverse transcribed into copia DNA. The present report describes the isolation of linear extrachromosomal copias whose structure closely resembles the analogous retrovirus provirus linears and whose synthesis is unaffected by inhibitors of the cellular DNA polymerase responsible for chromosomal DNA replication.

Animals↗
PubMedDate not supplied

Isolation of the closed circular form of the transposable element Tc1 in Caenorhabditis elegans.

The mobilization of Tc1, a transposable element in the genome of the roundworm Caenorhabditis elegans, has been investigated. Genomic blot hybridization has shown that Tc1 exists in very different numbers in the genomes of two closely related strains of C. elegans: there are approximately 30 copies of Tc1 in the Bristol strain, whereas in the Bergerac strain there are 200-300. Most of these Tc1 elements are structurally highly conserved although there exists a second form which contains a HindIII restriction site (Tc1 (Hin) form) and comprises approximately 10% of the population. Excision of Tc1 from its chromosomal location in the Bergerac strain is indicated by the presence, on genomic blots, of a minor band corresponding to the size of the uninserted restriction fragment. Here we describe the recovery of extrachromosomal linear and closed circular copies of Tc1 from the Bergerac strain, presumably a result of Tc1 excision.

Animals↗
PubMedDate not supplied

Rearrangement of nitrogen fixation genes during heterocyst differentiation in the cyanobacterium Anabaena.

Nitrogen fixation by the cyanobacterium Anabaena is carried out in heterocysts, specialized, non-dividing cells which differentiate under conditions of ammonia or nitrate deprivation. In Anabaena, heterocyst differentiation is accompanied by rearrangement of some nitrogen fixation genes. A site-specific recombination between an 11 base-pair direct repeat sequence flanking the nifK and nifD genes removes 11 kilobases of intervening DNA, resulting in juxtaposition of the two genes and an alteration of the nifD protein-coding sequence.

Cell Differentiation↗
PubMedDate not supplied

Bovine papillomavirus genome elicits skin tumours in transgenic mice.

Transmission of the bovine papillomavirus-1 (BPV-1) genome through the mouse germ line results in the heritable formation of fibropapillomas of the skin, a tissue-specific phenotype analogous to that observed in natural BPV-1 infection of cattle. Oncogenesis is slow, with tumours first arising at 8-9 months of age, usually in areas prone to wounding. Extrachromosomal BPV-1 DNA is detected in all tumours, whereas normal tissues show only integrated DNA.

Age Factors↗
PubMedDate not supplied

Novel submicroscopic extrachromosomal elements containing amplified genes in human cells.

In previous work, several methotrexate (MTX)-resistant variants were isolated frm the human cell line HeLa BU25, which exhibited a high degree of dihydrofolate (DHFR) gene amplification (estimated to be 250- to 300-fold). These variants did not contain any chromosome with a homogeneously staining region (HSR) and exhibited only a small average number of minute chromosomes per cell: these two types of karyotypic abnormalities generally accompany selective gene amplification. We now report that structures containing amplified DHFR genes in one of these variants (HeLa BU25-10B3) can be isolated by pulsed-field gradient or field-inversion gel electrophoresis as homogeneous DNA molecules of approximately 650 kilobases (kb). Electron microscopy of metaphase spreads from these cells reveals chromatin fibres with a similar DNA content, which are probably related to the above elements. These represent a novel type of extrachromosomal structures in mammalian cells.

Drug Resistance↗
PubMed1988

Maternal terminology.

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Extrachromosomal Inheritance↗
PubMed1970

Twin biology.

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Embryology↗