Methods for decreasing RNA polymerase III transcription in mammalian cells

Coppins, Beverly Ann (2007) Methods for decreasing RNA polymerase III transcription in mammalian cells. MSc(R) thesis, University of Glasgow.

Abstract

RNA polymerase III transcription has been found to be abnormally elevated in many types of transformed and tumour cells. These rapidly growing cells demonstrate an increase in pol III transcripts and hence, an accumulation of protein above normal levels. The accretion of protein may lead to uncontrolled cell growth, a hallmark of cancer. Thus, the ability of pol III to increase a cell's biosynthetic capacity links it with malignant growth. Therefore, experiments were conducted to assess whether pol III transcription could be regulated by decreasing transcription in mammalian cells. Homo sapien and Mus musculus cell lines were investigated to consider three different methods for decreasing pol III transcription; targeting a subunit of the transcription machinery, Brf1, by small interfering RNA, targeting the polymerase with the specific drug tagetitoxin, and inducing a negative effector of pol III transcription, Maf1. Levels of pol III transcripts were decreased in response to transfection of Brf1 siRNA, which also had a decreased effect on proliferation rates. When cells were treated with tagetitoxin during electroporation, pol III transcription also decreased. Induction of the pol III suppressor Maf1 decreased pol III transcripts to varying degrees, although faults were found within this system. More significant data was obtained by the application of siRNA against Maf1, as well from the analysis of samples acquired from heterozygous Maf1 cells. These results showed that decreasing Maf1 allows for the deregulation of pol III and an increase in pol III transcription.

• Item Type: Thesis (MSc(R))
• Qualification Level: Masters
• Keywords: Biochemistry
• Subjects: Q Science > QH Natural history > QH345 Biochemistry
• Colleges/Schools: College of Medical Veterinary and Life Sciences
• Supervisor's Name: White, Prof. Robert
• Date of Award: 2007
• Depositing User: Enlighten Team
• Unique ID: glathesis:2007-71015
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 09 May 2019 14:28
• Last Modified: 20 May 2021 20:05
• URI: https://theses.gla.ac.uk/id/eprint/71015