title: Reverse engineering the yeast RNR1 transcriptional control system.
abstract: Transcription is controlled by multi-protein complexes binding to short non-coding regions of genomic DNA. These complexes interact combinatorially. A major goal of modern biology is to provide simple models that predict this complex behavior. The yeast gene RNR1 is transcribed periodically during the cell cycle. Here, we present a pilot study to demonstrate a new method of deciphering the logic behind transcriptional regulation. We took regular samples from cell cycle synchronized cultures of Saccharomyces cerevisiae and extracted nuclear protein. We tested these samples to measure the amount of protein that bound to seven different 16 base pair sequences of DNA that have been previously identified as protein binding locations in the promoter of the RNR1 gene. These tests were performed using surface plasmon resonance. We found that the surface plasmon resonance signals showed significant variation throughout the cell cycle. We correlated the protein binding data with previously published mRNA expression data and interpreted this to show that transcription requires protein bound to a particular site and either five different sites or one additional sites. We conclude that this demonstrates the feasibility of this approach to decipher the combinatorial logic of transcription.
authors.affiliation: Department of Biomedical Engineering, Henry Samueli School of Engineering, University of California Irvine, Irvine, California, United States of America.
authors.forename: Grace
authors.initials: G
authors.lastname: Mao
authors.forename: James P
authors.initials: JP
authors.lastname: Brody
mesh_terms: D001665:Binding Sites; D002453:Cell Cycle / Q000235:genetics; D005240:Feasibility Studies; D015966:Gene Expression Regulation, Fungal; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D010865:Pilot Projects; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D048348:Reverse Transcription; D012264:Ribonucleotide Reductases / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D020349:Surface Plasmon Resonance
chemical_list: D009687:Nuclear Proteins; D009841:Oligonucleotides; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D012264:Ribonucleotide Reductases; C551024:Rnr1 protein, S cerevisiae