Aspects of in Vivo and in Vitro Transcription in Yeast

Yeast plasmids were employed to study the mechanisms of transcription by yeast RNA polymerase II and the yeast mitochondrial RNA polymerase both in vivo (in plasmid containing cells) and in vitro (in cell-free protein extracts). A complete in vivo transcription map of the yeast centromere plasmid YCp19 was determined, and the transcripts from the plasmid borne genes were quantitated. A plasmid position effect on gene expression was observed. Although YCp19 was maintained like a yeast minichromosome at approximately one plasmid per cell, several yeast genes overexpressed their RNA 3 to 5-fold above their chromosomal counterparts. The negative superhelical density ($-$d) was measured for yeast/E. coli shuttle plasmids isolated from yeast ($-$d = 0.02) and from E. coli ($-$d = 0.04). The superhelical density did not change when plasmids containing galactose inducible genes were induced to actively transcribe these genes. These results would favor models involving subtle DNA conformational changes to explain transcriptional activation.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biochemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Marczynski, Gregory Thadeus

• Chemistry, Biochemistry

Identifier

(UMI)AAI8823193

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/70570