title: Specific binding of a 47-kilodalton protein to the 3' untranslated region of rat transition protein 2 messenger ribonucleic acid.
abstract: The synthesis of transition protein (TNP) 2, one of the predominant nuclear proteins of mammalian spermatids, was shown to be posttranscriptionally regulated, by storing the untranslated mRNA for about 3-5 days in the cytoplasm of differentiating spermatids. It has been proposed that binding of a cytoplasmic protein to a conserved motif of 8 nucleotides (nt) in the 3' untranslated region (3'UTR) of TNP2 mRNA is involved in this translational control mechanism. In this report, we show that deletion or variation of the conserved 8-nt motif (GCCAT-CAC) in rat TNP2-3'UTR abolishes the capacity of the in vitro-transcribed RNA to reconstitute specific RNA-protein complexes in RNA bandshift assays. Using Northwestern analysis, we identified specific binding to the TNP2-3'UTR of four proteins of 45, 47, 49, and 60 kDa, all of which are stage-specifically regulated in male germ cell differentiation. Deletion of the 8-nt motif in the 3'UTR specifically prevented binding of the 47-kDa protein, the interaction of which is thought to be mediated by the RNA secondary structure. Analysis of the RNA secondary structure revealed that the 8-nt motif is an essential element of a specific stem-loop structure that is predicted for rat wild-type TNP2-3'UTR. Therefore we assume that the 47-kDa protein plays an important role in specific RNA-protein complex formation of rat TNP2-3'UTR that may be a central event in the translational control of rat TNP2 mRNA.
authors.affiliation: Institute of Human Genetics, University of Göttingen, Germany. mschlic@gwdg.de
authors.forename: M
authors.initials: M
authors.lastname: Schlicker
authors.forename: K
authors.initials: K
authors.lastname: Reim
authors.forename: G
authors.initials: G
authors.lastname: Schlüter
authors.forename: W
authors.initials: W
authors.lastname: Engel
mesh_terms: D000645:Ammonium Sulfate / Q000737:chemistry; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002868:Chromosomal Proteins, Non-Histone / Q000096:biosynthesis; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins; D005854:Germ Cells / Q000502:physiology; D008297:Male; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000096:biosynthesis*; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D014158:Transcription, Genetic / Q000502:physiology
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C489504:Tnp2 protein, rat; C053886:spermatid transition proteins; D000645:Ammonium Sulfate