E-GEOD-5290 - Transcription profiling of S. cerevisiae temperature sensitive eIF5A mutant shows accumulation of transcripts targeted to the Nonsense Mediated Decay pathway
Submitted on 11 July 2006, released on 13 June 2008, last updated on 27 March 2012
The highly conserved protein eIF5A found in archaea and all eucaryotes uniquely contains the posttranslationally formed amino acid hypusine. Despite being essential the functions of this protein and its modification remain unclear. To gain more insight into these functions temperature sensitive mutants of the human EIF5A1 were characterized in the yeast Saccharomyces cerevisiae. EIF5A is proposed to be an RNA binding protein. Thus, we used microarrays to identify special RNA subsets of up- and downregulation that are influenced by a limited eIF5A function in strains expressing a point mutated form of eIF5A. Experiment Overall Design: Wild-type and mutant yeast strains were grown at 25°C following total RNA extraction and hybridization on Affymetrix microarrays. cRNA probes derived from RNA preparations were obtained from four isogenic, haploid strains from each of the wild-type and the mutant. This ensured that changes in the transcriptome could be attributed to the impaired HYP function of V81G rather than to differences in the genetic background. Therefore, two groups of 4 datasets (per wild-type W303-WT and mutant W303-V81G strains) were generated after hybridisation to the micro arrays.
transcription profiling by array, unknown experiment type
Temperature-sensitive eIF5A mutant accumulates transcripts targeted to the nonsense-mediated decay pathway. Rainer Schrader, Craig Young, Detlef Kozian, Reinhard Hoffmann, Friedrich Lottspeich. J Biol Chem 281(46):35336-46 (2006)