Thesis (Ph.D.)--University of California, San Francisco, 2005. / Includes bibliographical references. Also available online.

Thesis (Ph. D.)--University of Chicago, Dept. of Molecular Genetics and Cell Biology, August 1997. / Includes bibliographical references. Also available on the Internet.

Dissertation (Ph.D.)--University of Toledo, 2009. / Typescript. "Submitted as partial fulfillment of the requirements for The Doctor of Philosophy in Biology." "A dissertation entitled"--at head of title. Title from title page of PDF document. Bibliography: p. 84-97.

Thesis (Ph.D.)--University of California, San Francisco, 2004. / Includes bibliographical references. Also available online.

Thesis (Ph. D.)--Ohio State University, 2007. / Full text release at OhioLINK's ETD Center delayed at author's request

Thesis (Ph. D.)--University of Texas at Austin, 2008. / Vita. Includes bibliographical references.

Transfer RNA genes require "positive" 5'-flanking sequences to direct efficient transcription. In order to delimit the modulatory sequences present in the 5'-flank of a Drosophila tRNA Val₄ gene, an extensive series of deletion mutants was constructed and end-points determined by dideoxy sequencing. The mutants were transcribed in vitro in a Drosophila Schneider II cell-free extract. Twenty nucleotides of the 5'-flank immediately adjacent to the mature tRNA coding sequence were required for transcription. Negative modulatory sequences were contained between positions -20 to -30 and -45 to -70 relative to the mature coding sequence. The -45 to -70 sequence shares homology with inhibitory sequences previously described in the 5'-flank of tRNA genes, except that this sequence was significantly larger in length. Sequences contained between positions -38 and -45 act as positive modulatory sequences which enhance the level of transcription. In addition, a Transcription Modulation Element (TME) was identified between nucleotides -33 and -38. The TME was also found in the 5'-flanking sequences of various other tRNA genes and preliminary data suggests that it enhances transcription efficiency through its position relative to the D and T control regions / Medicine, Faculty of / Medical Genetics, Department of / Graduate

Genetic variation within the cannabinoid 1 receptor (CB1R) locus (CNR1) has been repeatedly associated with drug addiction pathologies. Genomic annotation of CNR1 indicates the vast majority of this genetic variation likely results in altered transcriptional regulation of the CNR1 gene as a mechanistic link to the disease phenotype. There is a lack of information describing the regulation of CNR1 transcription and the potential impact of disease-associated variation within the CNR1 locus on its transcriptional regulation. This study investigates the impact of an evolutionary conserved regulatory region of CNR1, termed ECR1, and the disease-associated variation contained within, on the transcriptional activity of the cognate CNR1 promoter region. Reporter assays conducted in primary hippocampal cells demonstrate that CNR1 promoter exhibits variable transcriptional activity during periods of CB1R signalling and cell depolarisation. Coupled to allelic variants of ECR1, the CNR1 promoter shows significant changes in transcriptional activity under resting conditions indicating that disease-associated variation within ECR1 may decrease CNR1 transcription. Further, alleles of ECR1 can drive allele-specific transcriptional responses from the CNR1 promoter during periods of CB1R stimulation and cell depolarisation. The results highlight the potential for disease-associated regulatory variation of the CNR1 locus to create stratified transcriptional responses to specific cell signalling scenarios and putatively to clinical strategies employing pharmacological agents. Furthermore, investigation of DNA-protein interactions at the allelic ECR1 region demonstrate that disease-associated variation within ECR1 alters DNA-protein interactions within the nucleus consistent with a decrease in transcriptional activity in the disease-associated allele variant. Collectively the current work supports the hypothesis that disease-associated variation within the ECR1 regulatory region of the CNR1 locus has the capacity to significantly impact on CNR1 promoter transcriptional activity. It is posited that allele-specific transcriptional effects may have a major impact on the susceptibility of individuals to drug addiction or on responses to clinical pharmacological treatments.

published_or_final_version / Zoology / Doctoral / Doctor of Philosophy

published_or_final_version / abstract / Biochemistry / Doctoral / Doctor of Philosophy