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E-GEOD-8122 - Transcription profiling by array of fission yeast mutant for zfs1

Status
Released on 15 June 2008, last updated on 13 June 2013
Organism
Schizosaccharomyces pombe
Samples (8)
Array (1)
Protocols (5)
Description
Tristetraprolin is a vertebrate CCCH tandem zinc finger protein that can bind to and destabilize certain mRNAs containing AU-rich element binding sites. zfs1 is the single gene in the fission yeast, Schizosaccharomyces pombe, that encodes a protein containing the critical features of the tristetraprolin zinc finger domain. zfs1 has been linked to pheromone signal transduction control and to the coordination of mitosis, but no biological function has been ascribed to the zfs1 protein. Through a functional genomics approach we compared transcript levels in wild-type and zfs1-deficient S. pombe strains; those elevated in the zfs1-deficient strain were examined for the presence of potential tristetraprolin-like binding sites. One such potential target transcript was encoded by arz1, a gene encoding a protein of unknown function that contains armadillo repeats. arz1 mRNA decay was inhibited in the zfs1-deficient strain when it was expressed under the control of a thiamine-repressible promoter. Mutations within one AU-rich element present in the arz1 3prime-untranslated region protected this transcript from zfs1-promoted decay, whereas mutating another potential binding site had no effect. Binding assays confirmed a direct interaction between zfs1 and arz1 mRNA-based probes; this interaction was eliminated when key residues were mutated in either zfs1 zinc finger. zfs1 and its targets in S. pombe represent a useful model system for studies of zinc finger protein/AU-rich element interactions that result in mRNA decay. Experiment Overall Design: S. pombe wild-type and zfs1-deficient strains were grown to 1.0 A600 in EMM+5S at 30C for steady state analysis. Total cellular RNA was prepared from four replicate cultures.
Experiment types
transcription profiling by array, co-expression, genetic modification design
Contact
Citation
Characterization of zfs1 as an mRNA-binding and -destabilizing protein in Schizosaccharomyces pombe. Brandon J Cuthbertson, Yanhong Liao, Lutz Birnbaumer, Perry J Blackshear. , PMID:18042546
MIAME
PlatformsProtocolsVariablesProcessedRaw
Files
Investigation descriptionE-GEOD-8122.idf.txt
Sample and data relationshipE-GEOD-8122.sdrf.txt
Raw data (1)E-GEOD-8122.raw.1.zip
Processed data (1)E-GEOD-8122.processed.1.zip
Array designA-AFFY-47.adf.txt
Links