The Effects of TFEα and Promoter Sequence on Early Events in Archaeal Transcription Initiation

Expression of genes is dependent on promoter sequences in DNA, which direct transcription by RNA polymerase. Transcription factors interact with promoters and RNAP and are often conserved and are essential for proper gene expression. In this thesis, a functional region of archaeal TFEα, homologous to eukaryotic TFIIEα, has been identified, and specific amino acids in these regions have been determined to have significant importance in TFEα's ability to promote transcription in early initiation. All three Domains of life encode a 16S rRNA promoter, which is responsible for production of the 16S portion of ribosome. Transcription with this promoter is not well studied in Archaea but have been well studied in E. coli. It has been demonstrated that interactions with the essential, conserved transcription factor, TFB1, vary between gdhP and the 16S rRNA promoter from Pyrococcus furiosus. These experiments will provide a clearer understanding of the regulation of early transcription initiation, and identify critical differences between promoter sequences, as well as mutations in an essential transcription factor in P. furiosus that are required for function.