Caitlin McQueen

Turning on genes to build muscle

I received my BSc in Molecular Cell Biology from the University of York before starting my PhD Supervised by Dr Betsy Pownall and Professor Bob White the following October.

Transcriptional regulation during embryogenesis is necessary for correct cellular differentiation and function and a vast array of Polymerase II target genes have been identified as key developmental regulators. My co-supervision has allowed me to look at this transcriptional control more widely by investigating the role of RNA Polymerase III and its transcription of tRNAs during development. Recent advances in analysis of small RNAs and Protein-DNA interactions has enabled the exciting opportunity to study RNA Polymerase III regulation at genome-wide scale. This work has revealed a tightly regulated tRNA/mRNA interface in which RNA Polymerase III transcription matches the demand set by mRNA codon usage during development. This coordinate regulation indicates that transcriptional networks important for development are likely to be more complex than previously thought. RNA Polymerase III has 17 subunits only one of which has no equivalent subunit in RNA Polymerase II- Polr3G is unique to Polymerase III and exists in 2 distinct isoforms in mammals and in Xenopus. In mammals Polr3G is associated with proliferation and pluripotency, whilst Polr3gL is associated with differentiation. In order to characterise this tRNA/mRNA interface in more detail, I am analysing the regulation of Polr3G in vivo and Polymerase III transcription of tRNAs during myogenesis.

I am using CRISPR/Cas9 targeting, overexpression different RNA Polymerase III subunits and in vivo gene expression techniques to demonstrate the complementary expression profiles of Polr3G and Polr3gL during Xenopus tropicalis development, and the specific expression of Polr3G in the skeletal muscle lineage with a view to identifying their distinct roles. The transcriptomic analyses will be carried out using a novel microarray designed to investigate levels of specific tRNA transcripts under different conditions.