Characterization of zfs1 as an mRNA-binding and -destabilizing protein in Schizosaccharomyces pombe

J Biol Chem. 2008 Feb 1;283(5):2586-94. doi: 10.1074/jbc.M707154200. Epub 2007 Nov 27.

Abstract

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 3'-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.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • 3' Untranslated Regions
  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites / genetics
  • DNA Primers / genetics
  • Genes, Fungal
  • Models, Biological
  • Models, Molecular
  • Molecular Sequence Data
  • Mutation
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Phylogeny
  • Protein Binding
  • Protein Conformation
  • RNA Stability
  • RNA, Fungal / genetics
  • RNA, Fungal / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / metabolism*
  • Schizosaccharomyces pombe Proteins / chemistry
  • Schizosaccharomyces pombe Proteins / genetics
  • Schizosaccharomyces pombe Proteins / metabolism*
  • Sequence Homology, Amino Acid
  • Zinc Fingers / genetics

Substances

  • 3' Untranslated Regions
  • DNA Primers
  • Nuclear Proteins
  • RNA, Fungal
  • RNA, Messenger
  • Schizosaccharomyces pombe Proteins
  • Zfs1 protein, S pombe

Associated data

  • GEO/GSE8122