Replication of yeast DNA and novel chromosome formation in mouse cells.
To determine whether yeast DNA can replicate or segregate in mammalian cells, we have transferred genomic DNA from the yeast Saccharomyces cerevisiae into mouse cells. Most of the lines contained stably integrated yeast DNA. However, in two of the lines, the yeast DNA was maintained as numerous small extrachromosomal elements which were still present after 26 cell divisions in selection but which were lost rapidly out of selection. This indicates that, although yeast DNA can replicate in mouse cells, the yeast centromere does not function to give segregation. In one cell line we observed a large novel chromosome consisting almost entirely of yeast DNA. This chromosome segregates well and contains mouse centromeric minor satellite DNA and variable amounts of major satellite DNA which probably comprise the functional centromere. The yeast DNA in the novel chromosome has a compacted chromatin structure which may be responsible for the efficient formation of anaphase bridges. Furthermore, yeast DNA integrated into mouse chromosomes forms constrictions at the point of integration. These features have previously been presumed to be hallmarks of centromeric function in transfection assays aimed at identifying putative centromeric DNA. Hence our results suggest caution be exercised in the interpretation of such assays.
McGuigan
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Department of Biochemistry and Molecular Genetics, Imperial College School of Medicine at St Mary's, London, UK.
Huxley
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D000818:Animals; D001483:Base Sequence; D002503:Centromere; D018244:Chromosomes, Artificial, Yeast; D015825:Chromosomes, Fungal; D015342:DNA Probes; D004261:DNA Replication; D004271:DNA, Fungal / Q000096:biosynthesis*; D007739:L Cells; D008561:Membrane Fusion; D051379:Mice; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013104:Spheroplasts; D014170:Transformation, Genetic
D015342:DNA Probes; D004271:DNA, Fungal