Molecular and General Genetics MGG

, Volume 196, Issue 2, pp 323–331 | Cite as

Short nascent DNA pieces, accumulating in Saccharomyces cerevisiae after inhibition of DNA chain elongation, hybridize to specific chromosomal sites

  • Wolfgang Oertel
Article
Received:

Summary

In temperature-sensitive DNA chain elongation mutants (cdc8ts) of Saccharomyces cerevisiae even at the restrictive temperature a small portion of DNA is synthesized, which can be labeled by radioactivity. Under denaturing conditions this product sediments in alkaline sucrose gradients with about 4S. It is probable that these short nascent DNA pieces are derived predominantly from newly activated origins of replication. An alternative and more direct method of limiting the elongation of DNA chains uses araCMP as an inhibitor in nucleoside monophosphate-incorporating yeast strains. As with the cdc8ts mutants the only radioactive products in labeling experiments with [32P]dTMP are 4S pieces. Potential sites of their formation are small replication “bubbles” and terminal doublestranded loops observed in electron micrographs of the DNA from araCMP-inhibited cells. The pieces hybridize specifically with the replication origin of the 2 μm-plasmid, the chromosome telomeres and a group of chromosomal genes. Other genes and the centromere of chromosome 11 (CEN11) do not react. The 4S pieces hybridize with only three of nine cloned autonomously replicating sequences (ARS). It is concluded that ARS sequences, at least in the presence of araCMP, are not always used as replication origins within their normal environment on the chromosome.

Keywords

Saccharomyces Cerevisiae Nucleoside Sucrose Gradient Replication Origin Chromosomal Gene 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • Wolfgang Oertel
    • 1
  1. 1.Abteilung D (Membranbiochemie)Max-Planck-Institut für BiochemieMartinsriedFederal Republic of Germany

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