PhD (Doctor of Philosophy)
Date of Award
A uniquely human interspersed repetitive DNA sequence family, the L2Hs, are highly polymorphic in human genomes. Several features of interspersed repeated DNA may contribute to the instability observed. Certain motifs (direct repeats, palindromes, and inverted repeats) comprising L2Hs elements may adopt unusual secondary structures such as cruciforms or hairpins. These motifs have been associated with features of genome instability in recombination, insertions and deletions. The L2Hs elements also are AT-rich (76%) compared to the bulk of human DNA (52%). That their dynamic nature (i.e. polymorphisms) may arise from recombination, insertions and deletions has led to the hypothesis that the L2Hs element is intrinsically dynamic and may influence the stability of the surrounding genome. Thus, the stability of the L2Hs element was tested in a bacterial model system. A cloned 0.6 kb L2Hs element forms non-B-form structures in recombinant plasmids pN6 and pN2, which differ only in insert orientation. Instability of pN6 and pN2 plasmids was observed in serial propagation studies in which E. coli cells containing the plasmids were cultured every 24 hours for 28 days. The vector plasmid pTZ19U, as control, was found to be stable in all passages while the two L2Hs recombinants developed deletions of the L2Hs insert as well as adjacent vector sequences. The isolated deletion mutants have been characterized via restriction cleavage studies and sequencing to map the boundaries of the deletions. Direct repeats and potential stem-loop structures have been discovered at or within close proximity to the deletion boundaries. The data demonstrate that the L2Hs recombinants' unusual sequence features with potential for non-B-form secondary structures, influence genome stability via their involvement in generating errors during DNA replication and DNA repair.
Dissertation - unrestricted
Posey, Eugenia L., "The Influence of a Human Repetitive Dna on Genome Stability" (1998). Electronic Theses and Dissertations. Paper 2960. https://dc.etsu.edu/etd/2960