Understanding the mechanism of transcription of the genes that code for 45S pre-ribosomal RNA is essential if we are to understand both normal and abnormal growth processes, e.g. wound healing and neoplasia. Ribosome biogenesis, and therefore the expression of the ribosomal RNA genes, is coordinated with the rate of cell growth, and responds to a variety of signals, depending upon the cell type studied. The long-term objective of our research is to determine the mechanism(s) by which ribosomal RNA gene (rDNA) transcription is regulated. At least two trans-acting factors are required for accurate and efficient rDNA transcription, SL-1, and UBF. SL-1 is required for transcription while UBF activates transcription. Recent studies demonstrate that there are several ways in which rDNA transcription is regulated. One is the regulation of the ability of RNA polymerase I to initiate transcription. It has not been clear how many factors control the ability of pol I to initiate transcription. Two of the factors, Rrn3/TIF-IA and TFIC were believed to be the same factor. We demonstrated that they are two different factors. The yeast Rrn3 gene is essential for cell viability, and experiments in mammalian cells demonstrate that mammalian Rrn3 is essential for ribosomal gene transcription. However, there is considerable controversy concerning the role of Rrn3 in transcription, e.g. is it required for the recruitment of pol I to the rDNA promoter? Moreover, our data demonstrate fundamental differences between the mechanisms that regulate Rrn3 function in yeast and mammalian cells. Two of our goals focus on the determination of the role of Rrn3 in rDNA transcription and the mechanism(s) that regulate Rrn3 activity. Our third aim focuses on the role of PAF53, a second polymerase associated factor. Several lines of evidence suggest that PAF53 is an important component of the apparatus that regulates rDNA transcription. Antibodies to PAF53 block rDNA transcription, and the association of RNA polymerase I with PAF53 correlates with the rate of rDNA transcription. For example, PAF53 levels, but not core RNA polymerase I levels are reduced when NIH 3T3 cells are serum starved, while serum starvation causes the dissociation of PAF53 from RNA polymerase I in 3T6 cells. However, the role of PAF53 in rDNA transcription is yet to be defined.

R01GM069841

2005-09-01

2010-08-31