title: Functional equivalence of an evolutionarily conserved RNA binding module.
abstract: Members of the tristetraprolin (TTP) family of proteins participate in the regulation of mRNA turnover after initially binding to AU-rich elements in target mRNAs. Related proteins from most groups of eukaryotes contain a conserved tandem zinc finger (TZF) domain consisting of two closely spaced, similar CCCH zinc fingers that form the primary RNA binding domain. There is considerable sequence variation within the TZF domains from different family members within a single organism and from different organisms, raising questions about sequence-specific effects on RNA binding and decay promotion. We hypothesized that TZF domains from evolutionarily distant species are functionally interchangeable. The single family member expressed in the fission yeast Schizosaccharomyces pombe, Zfs1, promotes the turnover of several dozen transcripts, some of which are involved in cell-cell interactions. Using knockin techniques, we replaced the TZF domain of S. pombe Zfs1 with the equivalent domains from human TTP and the single family member proteins expressed in the silkworm Bombyx mori, the pathogenic yeast Candida guilliermondii, and the plant Chromolaena odorata. We found that the TZF domains from these widely disparate species could completely substitute for the native S. pombe TZF domain, as determined by measurement of target transcript levels and the flocculation phenotype characteristic of Zfs1 deletion. Recombinant TZF domain peptides from several of these species bound to an AU-rich RNA oligonucleotide with comparably high affinity. We conclude that the TZF domains from TTP family members in these evolutionarily widely divergent species are functionally interchangeable in mRNA binding and decay.
authors: From the Signal Transduction Laboratory and.
authors: Melissa L
authors: ML
authors: Wells
authors: From the Signal Transduction Laboratory and.
authors: Stephanie N
authors: SN
authors: Hicks
authors: Genome Integrity and Structural Biology Laboratory, NIEHS/National Institutes of Health, Research Triangle Park, North Carolina 27709 and.
authors: Lalith
authors: L
authors: Perera
authors: From the Signal Transduction Laboratory and Departments of Medicine and Biochemistry, Duke University, Medical Center, Durham, North Carolina 27710 black009@niehs.nih.gov.
authors: Perry J
authors: PJ
authors: Blackshear
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016880:Anisotropy; D019143:Evolution, Molecular; D017353:Gene Deletion; D055879:Gene Knock-In Techniques; D005816:Genetic Complementation Test; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry*; D010641:Phenotype; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry*; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D012568:Schizosaccharomyces / Q000737:chemistry*; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid; D051816:Tristetraprolin / Q000737:chemistry; D016335:Zinc Fingers
chemical_list: D009687:Nuclear Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D029702:Schizosaccharomyces pombe Proteins; D051816:Tristetraprolin; C097436:Zfs1 protein, S pombe; D012313:RNA
keywords: RNA binding protein; RNA turnover; mRNA decay; yeast genetics; zinc finger