Melanie J. Dobson

Structure and polymorphism of human telomere-associated DNA

We have analyzed the DNA sequences associated with four different human telomeres. Two are members of distinct repeated sequence families which are located mainly but not exclusively at telomeres. Two are unique in the genome, one deriving from the long arm telomere of chromosome 7 and the other from the pseudoautosomal telomere. One telomere-associated repeated sequence has a polymorphic distribution among the chromosome ends, being present at a different combination of ends in different individuals. These data thus identify a new source of human genetic variation and indicate that the canonical features of the organization of telomere-associated DNA are widely conserved in evolution.

Loss of 2 um DNA from Saccharomyces cerevisiae transformed with the chimaeric plasmid pJDB219

SummaryTwo plasmids containing Saccharomyces cerevisiae 2 µm DNA sequences and the S. cerevisiae LEU2 gene have been found to display incompatibility with 2 µm DNA; in the presence of the LEU2 plasmids, 2 µm DNA can be lost. The LEU2 plasmids can be lost spontaneously after (and before) 2 µm DNA loss has occurred, so that strains completely lacking 2 µm DNA sequences can be obtained routinely.

Relationship of the [psi] factor with other plasmids of Saccharomyces cerevisiae

Abstract The [psi] factor is an extrachromosomally inherited genetic determinant of the yeast Saccharomyces cerevisiae for which no autonomous physical determinant has yet been identified. Using both physical and genetical techniques we demonstrate that the [psi] determinant does not reside on other previously described yeast extrachromosomally inherited determinants; namely mitochondrial DNA, L double-stranded RNA, M double-stranded RNA, and [2 μm] DNA. Stable [psi+] strains, lacking one or other of these determinants were constructed. It is concluded that the [psi] factor may represent a new yeast plasmid.

Control of recombination within and between DNA plasmids of Saccharomyces cerevisiae.

Summary[2 μm+ and [2μm°] yeast were transformed to stable leucine prototrophy with the hybrid yeast — E. coli plasmid, pJDB219. This plasmid contains the entire sequence of the endogenous 2 μm yeast DNA plasmid in addition to the yeast nuclear LEU2+ gene and the Co1E1 derivative, pMB9. In the [2 μm+] transformants, a new wholly yeast LEU2+ plasmid, pYX, was generated, probably by a recombination event between pJDB219 and 2 μm DNA. The plamid, pYX, in the absence of 2 μm DNA, was found to exist in equimolar amounts of two forms, A and B, which probably arise by intramolecular recombination across the inverted repeat sequences of the 2 μm DNA portion of the plasmid. pJDB219 was found to require the presence of 2 μm DNA to undergo this intramolecular recombination. The results suggest that 2, μm DNA and pYX code for a gene product required in this recombination event which pJDB219 cannot produce.

Sequence variation in the LEU2 region of the Saccharomyces cerevisiae genome

The LEU2 regions present on yeast plasmid vectors come from two sources, a series of strains derived from S288c and strain M127. The LEU2 region from the S288c series contains a Tyl-17 element with its associated delta sequences and a small repetitive RNA gene while the LEU2 region from M127 which is present on pJDB248, lacks the Tyl-17 element, but carries a delta sequence and a small RNA gene. The various LEU2 plasmids currently in use vary with respect to these sequences depending on which restriction fragment from the region is present on the recombinant molecule. In addition, strain M127 contains three LEU2 homologous sequences that are represented by different EcoRI fragments and which segregate independently at meiosis. Therefore, there are at least four forms of the centromere-distal EcoRI fragment of the LEU2 locus in the Saccharomyces cerevisiae gene pool; these are 7.1 kb, 1.9 kb, 1.48 kb and 1.15 kb long.